OK, so I think we can start. So hello everybody. Happy to be here with you today.

So my name is Audrey Brunet. I'm the marketing and technical manager for Eminent for Central, Eastern Europe, and Middle East Africa. So, welcome to the second episode of our Berback Meal Quality minise mastitis diagnosis 2.0.

So for those who weren't with us for the first episode in March, so this ministry focused on the management of clinical mastitis and it is in a three part, the replay of the. First episode Mati is with the Boss new tool for no problem is also available. I will put after my introduction the link in the chat so that if you want to watch it after this episode, you can.

And for your information, the third episode will take place on September 28th so you can already write it on your calendar. Before we start to technical information, I would like to invite you to have a look at the chat room on the right side of your screen. So this chat room now is open and if you want to share some comments or any technical issue, it's there.

But for question, please ask them at the bottom of the screen. There is a Q&A box and, you can write your question there and we will answer them during the session or later if there are too many by email. In addition to that, during the session, we are going to ask you to participate to different pool and they will appear during the presentation.

Ricardo will mention it. You may, so yes, just to reexplain also for those who weren't with us the first time, you may ask yourself why we decide to develop this project. We are all aware in fact that populations are growing with more and more people to feed and the dairy products are one of the basis of our nutrition.

In addition, also to that consumer requirement in terms of quality, safety and traceability are increasing and at their level, part of our dairy strategy, we know that aca makes better meat. That's why we design in close collaboration with international experts to target 150 programmes to help dairy professionals for sustainable production of milk. This programme takes into account the main pillar of dairy farming, which are welfare, reproduction, nutrition, and milk quality based on the evaluation of key performance indicators.

And last time also I showed you that we have a Edited a book to support that. Today again we want to talk about milk quality reviews so because the effect of milk quality on production, animal welfare, and the farm finance makes it a major concern for most dairy farmers, vets, and farm advisers. The main disease on dairy farms remain mastitis, and it affects the efficiency of dairy farms and may negatively affect animal welfare and production as well.

So like I said, this is a mini in three parts focusing on milk quality, and the second episode will address the fundamental question relating to mastitis diagnosis in its multitude of layers. From the challenge of current clinical mastitis diagnosis in increasingly large form to the diagnosis of mastitis pathogen. It will cover the determination of new infection rates at different moments of the car production cycle and also address most of the information that can be obtained from milk recording beyond somatic cell count.

Finally, this training discussed possible sources of problems that may lead to increased total bacteria count in milk. Now I'm I'm very happy to introduce our speaker. So, the same as last time, it's the same for the next time also.

Ricardo Bexiga, he's in fact the author of the milk quality part of our target 150 manual. Ricardo has a huge experience in milk quality. He worked as a practitioner with a focus on dairy cattle for several years and worked in mastic diagnosis concurrently at the Faculty of Veterinary Medicine of Lisbon.

Ricardo was also a resident of the European College of Bovan Health Management in Glasgow, Scotland, and he was a diplomat of that college also since 2008, and he concluded his PhD on Bovin Mercedes also . At the Faculty of Veterinary Medicine in 2011, he's still working there, in fact, but he's also working in private practise at Cert, a practise with seven veterinarians working with Remina. He has also different publication in the international peer review, one book and a different chapter also in different book.

And last but not least, he's a form. President of the Portuguese Biatric Association, so very long, very long experience. Ricardo, and please if you have questions, do not hesitate to put them but on the Q&A box, and we will answer them during the session.

Thanks for your attention. Thank you, Audrey. Good afternoon to everybody.

I hope everybody can, hear me, well, loud and clear. So now we go into the 2nd episode regarding mastitis and milk quality. I hear that, you know, maybe, well, I'm hopeful that Netflix will pick this up sometime and do some sort of miniseries about it as well.

So, you know, millions can come pouring in. Anyway. I would like to talk about, the, the, the very, all the layers of mastitis diagnosis today and hopefully I can bring a little bit of, new information to, to, to you all attending.

OK, so first, I would like to try and see if We, we all can answer the surveys that we will have throughout the, the session by testing right now, what did you have for lunch is the, is the first question and I would like to see if you can all have access to, to the questions and, and supply an answer. So let's see if that's happening. I'm not sure if, if, .

Yes, the, probably we, when I send the, the que the list of questions, maybe I forgot the test question. Sorry for that. I think my understanding was that the, the, the questions were, were ready to, to go, I think.

Ricardo, they're in the chat box. Can you see the chat box? People are answering you.

OK. OK, so there are no. Now the pool, yes, I forgot to put this pool.

OK, so, so, so we'll we'll, we will not, . We will not have the, the, the, OK, so we'll we'll manage without that anyway, sorry for that, . So this was a little bit to try to increase the the interaction and to get a little understanding of what you're doing on your, on your farms and on your clients.

But, we'll, we'll have to do without that, OK? So, first thing I would like, to cover is really the challenge of mastitis detection. OK?

So we all know that managing mastitis is a very difficult task. It's a very costly disease that leads to very, Big losses in terms of milk quantity and milk quality, all the medicines that you have to pay extra in the extra labour, and also the extra investments on, you know, bedding materials, all the disinfectants you need to buy, for, for the prevention of, of mastis. So it's very, all in all, it's a very costly disease.

And we know that the early treatment of mastitis, clinical mastitis cases, is a key factor in the outcome and the cure rates of, of such cases. So it really we required to have a very early detection to boost our cure rates, so it's a determining factor, the early detection. It seems to be very simple to detect or it could be viewed as a very simple thing, the detection of clinical mastitis.

Clinical mastitis definition is changes in either the milk. The mammary glands and or the cow that are visible that are detectable on examination by the animal handler. Most often, the milker who's who's who's in close contact with the animal.

It seems fairly simple and straightforward, but actually it can be very difficult for, for a lot of people to agree on what is a clinical mastitis worthy of treatment. So for example, Would you say that this downer cow with mastitis is worthy of treatment? OK.

I would say, yeah, sure. It's oh, OK, so it's, it's working now. Yes, it's like I said, I forgot to put the question, but the other one, the others are here.

Very good. Excellent. So what do you think?

Is this clinical mastitis worthy of treatments? Just wait a few seconds and it'll come up on screen. The, the poll results.

OK, yeah. So, in, in, in, in such case, there's a downer cow with a toxic mastitis, probably if we don't treat her, we might, we likely lose her. What about this, this next situation here?

So there's an obvious other inflammation. What do you think? Is this worthy of treatment?

So, in this case, it's it's, it's fairly it's, it's quite interesting that a few of you are actually just saying no already, OK? But it's more or less the same distribution between yes and no. How about this next, next case?

Is this worthy of treatment or not? So, lots of clots, very altered milk secretion. Well, I guess it's no longer milk.

It's milk like, but. What do you think? Should this be treated or not?

So, 60/40. OK, so there's a more or less a pattern emerging that maybe the, the, the Even then these what might be seen as more or less clear cut cases, some of you are saying, I'm not sure if this should be treated. What about this, this situation?

Is this clinical mastitis worthy of treatment or not? Some of you, I'm sure are asking yourselves which is actually the mestitic sample? Is it the left or the right, you know?

This situation actually more clear cut split, so a lot of you are saying that this should be treated, OK? So it's a more serous-like secretion on the left, in the regular milk on, on the right. OK, but this goes to show you that, it's actually quite It's actually quite difficult for, for a panel of, of either farm managers or veterinarians to, to actually agree on which cases should or not be treated.

The very, well, I guess the most frequent case is a few clots in the beginning of milking, so there will be a few clots in the beginning. When you're first stripping the cow before milking, but then after milk let down, the milk is normal. Is this mastitis that should be treated or not?

So there's a high degree of subjectivity in, in when to decide to, to treat some, some cases, OK? And this is really made more difficult because sometimes there is no step in the milking routine. To eliminate the first squirts of milk to do the force stripping, OK.

So that makes mastitis, sorry, mastitis detection, even more difficult, and it's made even more difficult because The farms are increasing in size. Some farms are going from 2 to 3 milkings per day. There's a high pressure on farm managers to not increase labour costs too much, to not increase the energy that is is being .

Spent for, for a milking parlour to be working, sometimes, you know, nearly 2020 hours, 20 to 22 hours a day even. OK, so there's a lot of pressure and sometimes the option for the for the, for the farm manager is to reduce the force stripping to remove it from the, from the milking routine, OK? So some farms I go to, they've reduced it to only once a day, so 1 in 3 milkings per day.

Some are doing it 2 times a week, for example, and, and some other farms are abolishing all altogether, so they don't do the, the first stripping to detect mastitis. So what is the effect of that, of those practises on mastitis detection? Of course, it's probably going to decrease mastitis detection.

Unless we have ways of addressing this issue, certainly that is the case with automatic milking systems. So milking robot robots need to do this on a very regular basis. All the time they need to detect mastitis without human intervention.

So if they can do it with automated milking systems, we should be able to to do it as well. There needs to be, though, technology present on farm to allow for consistent and trustworthy detection of clinical mastitis with a reduction of human input, OK? And that is made possible in several different ways.

So the, the most, by far the most widespread and and longest, you know, that has been used in the longest period of time now is the electrical conductivity. This is based on the principle. That with inflammation, there is, there is increased vascular permeability to allow for the somatic cells, so, so the leukocytes to pass from the blood to the milk, .

And with those, with that increased permeability, what you'll have is also certain ions going through into the milk, which allow for an easier transmission conductance of electricity, of electrical currents in the milk. So when you have an increased electrical conductivity, you are, That's is translating the presence of an inflammatory response and hence mastitis. OK, this has been used for many years, but there are other possibilities.

That are being used mostly in in automated milking systems. So for example, looking to see if there is a different aspect, different look to the milk, that's, that's being used on some systems with the detection of colour changes, the sensors measuring either the reflection or the transmission of light generated by an LED for example. Some other system, so some other sys systems, sorry.

Are actually looking at measuring directly or indirectly the somatic cell counts, OK? So this is possible either through the formation of a gel, such as in the California mastitis tests. In this case, it's, of course, in, in, in systems, closed systems like this one shown on the picture.

Or they're actually based on staining of cells and counting these cells, OK? So this is again another possibility of an automated detection. On most automated milking systems, The detection is made by an index, which includes several of these indicators.

So you'll have an index, with a certain name, which includes, for example, a decrease in milk yields, an increase in electrical conductivity, and a decrease in the frequency of milking showing. Up as a sign that the cow that has these characteristics may have a mastitis and should deserve attention by the farmer. OK?

There are other possibilities. Sorry. Yeah, there is a question.

What the specificity of electrical conductivity? Yes, I, I, I don't know exactly. I could, I could look up for For papers for written on the subject, but I don't know off the top of my head what what would be the, the, the, the sensitivity or specificity for the detection of mastitis.

OK, I, yeah, you can look and then I can share with you. OK. Thank you.

OK, so, There are other, other systems. This of course is, is, is a little bit of a joke, but actually I know of a vet that tells me one of his clients, swears that he can detect subclinical mastitis by taste. And I'm not so sure if that is working or how the sensitivity and specificity are for that, but there's certainly other, other techniques, other, other technology that is being tested.

For example, the of thermal cameras or, or the measurement of the temperature of the milk have been used in the past to, to try to detect clinical mastitis with not a very high level of success actually. So it's not a proven technology and it's not being used massively, OK? So So it's, it's, it's, it's an interesting area for development as labour costs are, you know, increasingly important.

We might see different technology coming to be used more frequently on farm to detect clinical mastitis, . You know, more rapidly and allowing for a more rapid assessments of of the case and treatment, OK? So the other aspect, another aspect of Of mastitis is is actually what pathogens are causing mastitis on, on, on our farm.

And I would say that the first approach could be through the use of the boat tank PCR, which is an excellent tool for this, for this, first approach to a farm that we don't know yet, and that actually should be then performed. At least once a year to monitor to see if the if the situation is not changing. It is fairly cheap.

It shouldn't cost more than about €30 to do this PCR, depending on where you are, but €30 is more or less the the going price for, for. For, for this PCR, it's representative of the whole lactating herds and it has an added advantage because it can be used on farms that are not your own if you're sourcing some replacement heifers, for example, from, or cows from another farm. So, OK, you can see what's, what actual pathogens, mastitis pathogens exist on the other farm before you buy animals from that farm.

OK? So another, another question for you. Have you ever used the PCR test on on both tank milk?

So we have different possibilities, mastitis. Parasitism or reproduction. OK, so, Overwhelmingly, I guess you're saying that mostly you have not used the PCR test on your bulk tank milk.

Some of you have used it for reproduction, maybe see if there is DVD IBR, you know, Neosporra, other, other agents. For parasitism, I guess mostly people use it for, for, at least, well, fasciola, so fluke. And for example, for Otaragia also, especially, of course, in, in countries where there is some degree of of of pasture-based feeding, OK.

For mastitis, very low number, but it's actually quite a useful tool, . To use, it's, like I said, it's cheap. It gives you information about many different pathogens, OK?

The most widespread PCR commercially available allows for the simultaneous detection of 15 mastitis pathogens and one antibiotic resistance gene. In this case is, is penicillin resistance gene. OK.

So this is, I apologise, I, this is, for example, one done in Portugal, so it is in Portuguese, but you can check out the, the different, different pathogens. This is, this IAC is an internal control, and this is, of course, positive and negative, and you can check out the different pathogens that you can check for. OK.

I would recommend using this as a single pool of 3 samples. OK. So, if I'm going out to a farm, I'll ask the farmer.

To check or to collect a sample in 1 day, then 3 or 4 days later and to freeze these samples, and then I'll go over on my visit to collect another sample, so I have a sample from 3 days which I pulled together and send it out to the lab. This way, you increase the chances of, of Of, of having more pathogens detected because the excretion of some of these pathogens is intermittent. It's not continuous.

So, you increase the chances of, of picking up some of these, some, some more of these pathogens. Like I said, I would do this once a year as a control, if and more of that if the farm is open, so it's if it's buying in replacements, OK? It, I would say it's especially interesting, if you use it to focus or to detect Staphhorius, streptococcus yalactiae and mycoplasma bovis, which, in the past or even now, are viewed as contagious, .

Pathogens. So certainly, for some of them, especially for Streptococcus agalactia, it does not live outside the, the, the other. So it's necessarily being transmitted from cow to cow, in the milking parlour.

Stephorius, OK, might come up from the environment every once in a while. Mycoplasma bovis. It can, can come up from the, from the coughs as well if they were drinking contaminated milk as well.

If, if, you would be also looking to see if you find Prototheca, which is an algae, of course, it's not a bacteria. The cure rate of, of, of prototheca mastitis is very low. You cannot, of course, treat it with antibiotics.

So it, I find it also very interesting to, to, to detect in these in these cases, OK, in this, in this, in this panel. Some of the other pathogens, because they are, they can exist in the environment, you're not 100% sure if they're actually coming from a faecal, for example, contamination of the mammary gland or if they're coming from, from mastitis from the, from the others of the cows, OK? As I said before, These, these agents that I mentioned traditionally were were viewed as contagious and they still are, but nowadays, the, the frontier between contagious and environmental pathogens is, is blurred.

It's not so clear cut anymore. And that is especially so for strepto. Caucus uterus, which was viewed in the past as as mostly being an environmental pathogen, but which now we know that it can be, it can be working as an environmental or a contagious pathogen.

OK. So it's being both transmitted in the milking parlour and acquired from the cow's environment. This is very, very well illustrated by this, this, this study which was done in the UK involving 52 farms which were followed for 12 months.

In which 63% of the, the farms showed shared strains between cases of clinical mastitis. So there was on 63% of the farms, there was evidence that there was being cow to cow transmission in the milking part. And in such farms, about 40% of the cases showed evidence of such transmission.

So in the, in the black, you see the cows in which there was, you know, cow to cow transmission, and the mastitis caused by the same strain, and then the grey, you'll see the . The cows in or the clinical cases of mastitis in which there was a you know, the, the, the strep uber was being acquired from the environment. OK?

So, like I said, it's, it's the frontier is not so clear and that can be the same. For even E. Coli or other pathogens that can be transmitted in the milking parlour between between cows, of course.

What can we do if that happens, if we have evidence of Stephhoria, Streptococcus galactia or mycoplasma bovis in the herd. Well, we can identify the positive cases, this might be, you know, More or less difficult, we'll talk a little bit about that. We can separate the cows and milk them last, or we can cull them if, if need be, or we can treat them.

For example, streptococcus acylacty has very high cure rates and it will cure if you do, if you do antibiotic treatment in most cases, OK? Another, another recommendation I would, I would I would follow would be to start using some type of cluster spraying system. With the parasitic acid at 0.03%.

It will decrease the, the life of the, of the rubber or the silicon, but, it will allow for a break in the transmission and the contagious in most cases, from cow to cow. So what if you're looking, what if, yes, yes, a question regarding what you just said regarding the disinfection of the cluster. How much time does it take to, to do it and we need not make the milking rating, too slow.

Yes, it will make it, slightly slower. Not a lot. From my experience, these, these systems can be integrated in a normal milking routine and maybe, the last one I put in is about 300, 350 cows milking.

It takes them after a whole milking, which is about. 2 hours and 45 minutes. It takes them 5 extra minutes at the end, not more than that.

So it's, it's, I've, I've, I've, I've had it also in, with goats. As well, with contagious problems, it's, it takes a little bit more time because the milking routine there is, it's almost nonexistent. So it, it does a little affect a little bit more if we're talking about goats and if you're talking about, cows, but it certainly does not add a lot of time to, to, to, to the milking.

Another aspect is, normally if there are contagious pathogens. One farm, one thing I do is stop for stripping, OK? So I tell them, do not force strip because the hands of the milkers are adding to the risk of transmission of pathogens between cows.

So certainly, if they stop for stripping and add this, they'll save time at the end of the milking, OK? Of course, there may, you, you might say that there are also problems with the stopping for stripping like we saw before, but I think if there is contagious pathogens circulating on the farm, that's something you, you need to do. OK, and still on the question, this infection of the cluster, what should be the frequent of spraying?

How many time I do, I, I, I recommend it every upon every cow exiting the, the, the, the milking parlour. OK, so once the cow leaves, spray the, the four clusters and, and they're gone and And what is enough. Yeah, it's, it's very, it takes, you know, spraying the clusters takes.

3 seconds, 4 seconds maximum. It's, it's very potent. The, the, the parasitic acid is, is very, it's a very potent, disinfectant.

It, it works in the presence of milk, but it's, it's very rapidly degraded. So it's, it, it is, . A food grades, disinfectant, if, if there is such a thing, if it goes on to the milk, it just, it just, is degraded.

So there's no issue with being, with a little bit of going into the milk, for example, if, if, if there is an accident, because it will just disappear. . What I see is that it does reduce the life of the rubber, OK?

And if it's sprayed onto the eyes, so some farms start using glasses on milkers because of that, but I mean, you don't really need to if you're careful. OK, and then in terms of the rubber, life, you would say that what is the normal duration without this step and we talk about 2500 milkings for for the, for the rubber. But normally, farmers do, do a little bit more.

They go a little bit more than that. I would say if you're using this, you, you would definitely need to, to, to respect the 2500 milkings. Mm, OK.

It, it's, it the, the consumption of the, of the acid is very low as well because you're using it on a very, very low percentage, you know, a very low concentration. And the system, by the way, I, there's different systems. This system that is being shown here.

To, it costs about €600 to have it for a, for a whole, for a milking parlour. For example, a 40 point milking parlour, it costs about €600 at least, it costs here in Portugal. It's a British, it's a British band brand, by the way.

OK. OK. Thank you.

Right. So, So going, going back to to what I was talking about, the detection of these pathogens. OK, if you're looking to detect mycoplasma bovis in individual cows, it could be a little bit tricky because not every, every lab does, the, the simple bacterio or the conventional bacteriology for mycoplasma bovis.

It's difficult to do and not a lot of labs can do it. The PCR in this case, if you're talking about individual animals, like I said, a normal PCR costs around €30. If it's even if it's just for mycoplasma, so it's expensive for a single animals.

So if there is mycoplasma bovis in the in and if you're looking to detect it in individual cows, you will spend a lot of money probably to to detect these cases. It's not such a case with Staph aureus, you can use This this culture media, which is called Manito salt agar, essentially, it has a high level of of salt, so most pathogens do not grow in it, but Staph Staphylococci will grow and Staph aureus will change the colour of the medium to to yellow, OK? It might occur with other staffs that it changes to yellow, but certainly you can screen a whole herd, with the, with a very, very cheaply because these plates will cost less than 1 year in the vicinity of 50 cents in most places.

So you can detect Staph aureus in a cow and you can do a whole herd testing if, if you want it fairly cheaply, OK? So what samples should you be sending for bacteriology? I would say probably the very high cell counts because they will, if you're doing.

If you're looking at what is causing mastitis on your farm and you're not focusing only on the, on the clinical mastitis. I would look for the very high cell counts because these, of course, have a bigger impact on the cell count in the bulk tank, but also look at chronically high cell counts. So maybe you're looking at above 500,000 cells, but for months, over 4 months or more.

So there's a chronicity here, and you need to consider also that for some agents, namely for Staphhorius, the cell count. Resulting from an infection of Stephforis might be fairly low. So sometimes you'll have a cow that has an average of 200,000 cells per mL and it is infected with, with Staphhoria.

So you need to be careful with those situations as well. And that's because One of the ways that Staph aureus has got to evade the host defences and also the antibiotics that are thrown at him, at them. Is the internalisation into cells, either epithelial cells or leukocytes.

So, Staph aureus will internalise. Cells and survive inside being excreted not constantly and actually causing a low response by the host by the cow, OK? This could happen with other pathogens.

Strep hurus can do this, but they will always cause a higher amount of inflammatory reaction, OK? And that's one of the reasons. Why both Staph aureus and strep uber actually have lower cure rates than most other mastitis pathogens, and they actually benefit from extending the antibiotic therapy for a longer period of time.

So, for example, if you have A mastitis treatment protocol, which is 4 days long, for example, you might need to double it to 8 days if you're facing Stephorus or strep huris to have the equivalent cure rates, OK? Just, just a little bit, I would like to talk a little bit about the, the, the, also the, the, the treatments of grade 1 and grade 2. So grade 1 being caused with changes in the milk alone, grade 2 in the milk and in the mammary gland, in, in, in.

In cases of, of grade 1 and grade 2 clinical mastitis caused by gram-negative bacteria, the use of antibiotics or it's non-use leads to the same cure rates. So essentially, if you're having mastitis, mild or moderate mastitis which is being caused by gram-negative bacteria, where you're no better off if you use antibiotics, OK? And that is why It is so interesting to use on-farm culture.

On-farm culture is, is primarily used to support the treatment decisions around the, the, the mild and moderate clinical mastitis to distinguish between. You know, no growth, so there's no bacteria, that is actually causing that mastitis or gramme negative, in which case you, you don't need antibiotics or gramme positive, in which case you will need antibiotics to treat, to treat that case. And also, like I said before, it's useful to identify Stephlois and strep ubris because in these cases, you'll, you'll have to treat for a longer period of time.

So I'm curious to, to, to know if if you're using or not these on-farm tests to To the, to, you know, to know what mastitis pathogens are causing a certain mastitis. So use on farm. And in the meantime, there was also a question with what you mentioned when you were talking about PCR.

I'm not really sure. If we applicate PCR, does it have an interference with other pathogen or negative fold or positive false positive or? Yeah, I'm not really sure about the question.

What you want to mean maybe, do you see? Yeah, I, I'm not, I'm not sure. I can see, I can check the question.

Yeah, because I'm trying to, to make it . But I'm not OK. I'm not sure if I understand, but .

If, if you're using PCR. You need to be careful on, on how you collect the sample, of course, because some, some people are using it, directly from the milk recording. Cups?

So in this case, of course, if a cow is there, before and it has a certain pathogen, it leaves, even though if you rinse it off, you, you, you still might find it over the next cow. So you need to collect the sample aseptically, so with all the procedures for, for normal bacteriology. .

You, you can use PCR which is only for mycoplasma or mycoplasma bovis. So without interference from other, from other pathogens like the other PCR, it's a multiplex PCR. It looks at many bugs, but in this case, you can use a PCR only for mycoplasma.

. And it has another difference, because being a PCR, it will also detect. Dead bacteria. OK?

So it, it, it doesn't tell you if the infection is active because if you have dead bacteria, the PCR is looking for the genetic material. So for that, that bacteria, it will still show up as a positive result. OK?

So I'm not sure if I, if I managed to answer, but those would be my, my main concerns about the use of PCR. OK. We, we can get back to that if in the end, if, if, if need be.

So the most frequent answer, yes, sometimes, I would say. And some, some, some people are not the second most frequent, no never, so I would encourage people that have not, used it in the past to start using it regularly, because it's an excellent tool. It allows for, big savings and certainly the farmers.

That we've started that we recommended and helped implement the system on, on farm. We've seen . If you know, they, they, they, they enjoy it a lot because they see that they can save a lot on milk and a lot on antibiotics as well, even though, quite often they will start using more non-steroidal anti-inflammatory drugs, OK?

There are several systems available, so. These are the three that I see most, especially the 1st 2 that I see most frequently being used in, in Portugal, and it's, it's, and it's quite, these two, the first two are, are, are being used quite often on, on, on bigger dairy farms, certainly in the south of Portugal. There are some issues, about sensitivity and specificity, but you need to consider the different objectives.

So I would, I would mostly use it to say there is growth, there is no growth, and if there is growth, it's, it's gramme positive or gramme negative. OK, if, if you're trying to look to identify a specific, you know, species of bacteria, then you certainly your sensitivity and specificity are going, are going to drop, quite a lot, OK? Also, if you start if you're kind of too demanding, with the farmer, OK, you can detect this and that and the other, certainly, for example, as, as in this scheme and this type of plate.

Which is also available on the market. It can be too complex and that it can discourage the farmer or the farm manager from, from using it, OK? 11 point which I find is, is difficult to manage is, is that a lot of yeasts.

Grow in 24 hours and they look the same as staphs. OK? So sometimes you're treating because you think it's staphs and it's actually yeasts, and that's the most frequent, situation which I, which I find on farm that, throws people off using this system more, more frequently.

OK. We also have Verback has, has got the, the, the system, the speed map colour. Which, which I've been using as well.

It's, it's got several interesting features to it. It can identify, up to 8 pathogens, including, and this is quite interesting, the mycoplasma. OK, so, If you have a, a, a, a, a, a bulk tank PCR which is positive for mycoplasma, this would be a possible solution for you to actually detect the cows that are positive, OK?

Because there's not a lot of options in the market for you to detect mycoplasma. . It also has the advantage of, of, of allowing for antimicrobial susceptibility testing for 14 different antibiotics or antibiotic combinations.

OK. So it allows you to, to not just perform the identification, but also check if there are antimicrobial resistance specific to, on your farm that allows you to, to you, to be more targeted in your approach, OK? So in this case, it's, it's a biochemical identification, gallery, like, The API system, which is some of you might be familiar, which, which is used on, on a, a lab environment, not on farm, mostly, but this, in this case, you can, you can do it on farms if you have, if you have an incubator, OK?

It, it allows you to read the susceptibility testing results in 24 hours and allows you to, to To know the, the identification in 48 hours for, for all the pathogens except for mycoplasma, because that will take 7 days, OK? So, normally in a, in a, in a lab environment, mycoplasma, you can, you can read the result after 10 days. In this case, a little bit earlier than that, but it still will take 7 days because it's, it's, it's a very Fastidious microorganism takes very long time to, to grow, OK?

OK, so I would like to change a little bit about about sorry, change the direction of the, of the diagnosis now into the new infections, OK? So, In fact, before you start, sorry, I have two questions for you. One that was linked to what you just said talk about the on-farm culture, that some veterinarian may see the use of on-farm culture as being a task for the vetter, not for the farmer.

What is your opinion on this? OK, so, so I guess it depends where you are and, and, and what kind of, what kind of client you have. Certainly I see it, even in Portugal, it's being used in different in different ways on, on, on some farms, smaller farms.

It's with less technical, less technically developed farms, I guess. It's the vet that does this, so you'll have an incubator and it pro it provides this service to to a farm or to a farmer that might not be able to do it on his own. Certainly for bigger farms.

And as the ones that we have here in the south of Portugal for 400, 500 cows, it's something that the, the farmer can do. He normally has a good technical knowledge of, of, and he can be trained to, to do this, and there's still a role for the vet, because, you know, the vet is the one that implements the system. That designs the protocols, the treatment protocols that assesses if they're working or not.

And it's, it's the vet that will get a picture of the plates. I get very often in the beginning, I get pictures sent to me via WhatsApp message and they'll, they'll ask, OK, what's, what is this? What should I do in, in this case?

Is this, should I treat like this or like that? For example, if there is growth in all, in all quadrants of an all culture media, it might just be a, a, a contamination. What should they do?

So there's still a role for, for, for the veterinarian to participate in, in aids with using such systems. So my answer, in short, it, it depends what kind of, what kind of client or what kind of farm you're facing. OK, so smaller, less technical, it's for the vet, I guess.

If they're more technical, I would, and bigger, I would use it directly on farm, no problem. OK, and the second question is, in fact a good transition I guess with what you are going to talk after. And there was a question regarding at summertime clinical mastitis are more frequent, how to manage to reduce its impact on milk quality.

OK. Yes, excellent questions. I guess it, it depends where, where you are in the world, because, summertime for me, it might be different from summertime from somebody else, you know, Summertime for me is, some days of 40 °C, lots of cooling, lots of water going on on the cows, which, you know, Dragging, dragging dirt on cow's teeth sometimes.

So it's, it's, it's a challenge, but it, I guess it depends, depends where you are in, in the world and, and, and what kind of conditions. From, from my, from my side of things. I see that the, the, the decrease in feed intake might affect the immunity of the cows, the excessive water, in some types of, some types of cooling systems.

Also needs to be controlled so that the water cannot drip from, from the cow's teats. But certainly you need to cool the cows, otherwise there will be, you know, a decrease in fertility, a decrease in milk production, a decrease in feed intake, a decrease in, in, in animal welfare. So, so, but like I said, it depends on where you are and what kind of problems you, you might have on a specific farm.

Mm. Not sure if I answer that. Maybe I could, if I knew a bit more about some cases I could, I could maybe elaborate on a little bit more, more on that.

OK. So, I'll, I'll, I'll, I think I need to speed up a little bit because I'm conscious of the time and of the things I need to talk about still, OK? So I'll, I'll, I'll try to speed up a little bit of things.

So one thing that you need to, to also do in terms of diagnosis. I know how many new infections are occurring each month. So you need to calculate your subclinical mastitis monthly incidence rates, which is the number of cows from all the, the cows in at risk.

Which have gone from below to above 200,000 cells per, per mL per mL of milk, OK. Sometimes this is, this is available directly from the milk recording company. Sometimes you need to calculate it yourself by using the, the, the threshold of the 200,000 cells, OK?

So if you're going above that, there's a new infection. If you're lowering from above to below, there's a cure. If you're in two successive months you have a cell count above 200,000 cells.

It's a chronic case, case, and if you're below in both months, then it's a healthy, it's a healthy cow, OK? It is a very good measure on of how your preventative measures are in place for mastitis. So if you're doing a good job of preventing mastitis, this will stay.

In in the fairly low and your target should be about 5% of cases per month. I, I see from my experience, if you're below or around 7%, normally there is a decrease in somatic cell count average. So certainly you want to be at 7 or below 7%.

For your cell count average not to increase, every month, OK? The target for the chronic group is to stay under 10% as well. If you have a bigger proportion than these 10%, that may be indicative of problems with, with pathogens with a low cure rates for, for example, staphorus, OK?

So if you're looking at at the manual, the T150 manual, that this is all very well explained, or hopefully well explained in in in the manual, and we use this system of, of, of with the colours to, to, to indicate what kind of situation you have on your, on your herd on on your client. Another, another important Piece of diagnosis in, in, in mastitis is diagnosing what is actually occurring during the dry period, OK? How many new infections are occurring in the dry period and how many infections are actually being cured by the protocol you're using.

Again, what you're going to do is to use the 200,000 cells per mL threshold to define infection, and then you're looking at the, the comparing. The last milk recording before drying off, and the first milk recording after calving. So you're comparing those two, and you're seeing, you're seeing what kind of dynamic exists in terms of somatic cells, OK?

So if you, if you were below 200,000 cells and increased to above, that's a new infection. You want that to stay under 10% of all the cows that were not infected at dry off. If you're looking at.

A cow that had a high cell count above 200,000, and, and after calving it was below, that is a cured cow, and really you want to be at over 85% of the cows that were infected at dry off. You want them to be cured after calving, OK? Then of course you have the correspondent not cured and healthy cows, of course.

Even without looking at these numbers, you might get an idea of, of a problem in the dry periods or transition or transition. Beginning of lactation, for example, if the average cell count for the cows in the beginning of lactation is the highest in all the cows that are being milked on your farm. OK, so, for example, this is a direct translation of the milk recording, one of the milk recording charts that I have here in Portugal.

It gives you, it splits the herd into cows being in the 1st 60 days. After calving on the 61 to 120 periods, etc. And certainly for a cow like this, if I for, not for a cow for a herd, sorry, if you see a herds in which the highest cell count comes in the beginning of lactation.

That certainly means that there is an issue in the transition periods and the cows, even though they had antibiotics, even though they had maybe teat sealants, they're still showing that they have a highest, the highest cell count in the beginning of lactation. OK. Again, something from the T150 manual.

You want really your dry period cure rate to be above 85%. So all the cows that were infected at dry off, you want over 85% of those to be cured after calving. If that's not occurring, maybe you have some difficult pathogen causing mastitis.

Maybe you need to revise the dry off protocols to see if, if that improves, OK? So this is just An example of of what I get and and and and don't worry, I'll not show this in Portuguese. I have little little bits translated into English, but this is what a summary of the milk recording in in Portugal for for a particular farm.

OK, this is, this is, for example, this is a a a herd with 500 cows. And I've translated bits of this. This is, for example, another list we get is the list of the problem cows, all the cows that are above 200,000 cells per per mL, and you get the dais and milk, the somatic cell, the linear score, which is the logarithmic trans .

Transformation, the, the, we also have the number of times that in the present lactation in which the cell count was above 200,000 cells. So you have a lot of information that might allow you to target your bacteriology that you're performing on on farm or making, or maybe even making calling decisions later on, OK? So looking at all the information that milk recording tell us, one such information is the proportion of heifers and the in, in the lactating herds of your farm.

So I would like to ask now, what, what do you know is the proportion of heifers. And the lactating herds, so the cows that are lactating on your farm. So some of you will not be farmers.

So you might not get a lot of answers, but anyway, . We see here that the most frequent answer is 30 to 40%. It would be the same for me on most farms that I go to is between 30 and 40%, and some, a lot of, a lot of answers actually are below 30%.

OK? So this is, this is, if you're looking at a milk recording at the snapshots for the farm for a certain month. We can actually look at the proportion of heifers in the lactating herds.

So for example, here in these two cases, these two farms, we have On one, about 50% of the lactating hers is heifers. On the other 31.6% are heifers.

So I would say that if I was going to be a cow, I'd rather be in the second farm because maybe their culling is lower, OK? One of the reasons why you might have a big part of a bigger part of the population of the lactating herd, being in the first lactation might be due to excessive culling, OK? If there is, if, if it is such a case, where you need to investigate the culling causes.

Why, why is there excessive culling? OK. Why, why is it?

Is it mastitis problems, that is, is it fertility problems, OK. Of course, you might Be trying to increase the herd size, OK? You've just made a new housing for cows and you're trying to get it full.

So you actually have more heifers, and you're actually maybe buying in animals to, to, to fill the new, the new cow. Housing, the new shed, OK? In which case, you need to be careful that you don't run into biosecurity problems.

So for example, if you're sourcing out your heifers, maybe you want to do to check if they have BVD, IVR, etc. But also You want to see if they have mastitis pathogens by pathogens by doing a bulk tank PCR on the farm of origin, for example, OK? Another question I, another question I would like to ask you is, what is your lowest average days in milk for the herds?

OK, so the lactating herds, what is the average? Days in milk at the lowest time of year when it goes the lowest. OK, so, so split between 150 to 170, 170 to 2 100 days.

I would, I would be in the same situation, most frequently, OK? So it's, it's on most farms. So of course, you all know, it's a very, again, here we have a couple of real, real situations and the average days.

In milk for the herd in this particular month was quite different between farm A with 237 to farm B, which has about 160 days in in in milk. So this is a great a great measure of the reproductive efficiency of the farm. If there is Yes, sorry.

There was a question regarding, the excessive culling, can a farmer get into trouble by the veterinian authority or from by the authority or because of excessive, culling? I guess it depends on where you are. Certainly.

In in Portugal, no, but they would probably get in trouble, by some certifying bodies, OK, so. So the industry that is buying the milk, the dairy industry, or the supermarket chain that is buying the milk, might have, such as the case in Portugal at least, it might have an animal welfare, scheme, for them to certify that the cows on a particular farm that is, you know, From which their milk is coming from, has a higher higher welfare standards, and they will be looking at those, at those indicators, and also, you know, different things like flight, flight distance from the, the, the person assessing or I don't know if in some countries you might actually get in trouble from from a government or authority, but certainly from a a a a company that is buying the milk, I, I would say that could be the case, yes. Yeah So I was, I was, I was talking about the average days in milk for the farm being a good measure of reproductive efficiency.

If, if everything's going according to plan, if the cows are getting in calf fairly soon, then you should have a a a a low. A low number, average days in milk, of course. If you have.

If you have, some kind of, some level of heat stress in the summer time, the conception rate will decrease, over the summer, and normally you have some Even though it's unwanted, you'll have some seasonality in carvings. So, for example, in Portugal, I know that in the autumn, I'll have lots of carvings going on from September to December. I'll have most of the carvings going on on a certain farms, so in January and February, I'll have the lowest average days in milk.

Which tend to be around 150 to 1160 through, and then, as the year goes on, it might go up to 180, 190 for a, for a good farm. OK. So this is normal variation due to unwanted seasonality with the heat stress, but that's, it's, it's, this is a situation, for example, in, in my latitudes.

OK, you can also look at indicators from for, for a negative energy balance. OK, you've seen this before, this is the split of the herds. Based on days in milk, and if, if you see here in this particular farm, in the beginning of lactation and even up to 4 months after calving, the, the protein percentage in milk, the average protein percentage in milk is quite low, it's below 3%, which is an indicator of negative energy balance.

So, normally cows in negative energy balance will show . Different things, a milk protein below 3.05%, you should have really less than 15% of the cows in this situation.

By the way, this is, this is from an excellent review paper by Finbar Mulligan. He's one of the co-authors of the T150 manual, and this is a very, very, very good review about these indicators. In the transition period.

OK, so also the fat to protein ratio in milk being above 1.5, that's also an indicator of negative energy balance, and then having milk fats over 4.8%, which is an indicator of some level of of of fatty liver, also and and and indirectly an indicator of, of negative energy balance and the beginning of lactation.

This is very influential. In the transition periods, because of its impact on metabolic disease and on the immune competence of the cows. So if you see severe negative energy balance or occurring in most cows, you'll definitely know that you'll be having more metabolic disease and more infectious disease because the immune competence of the animals is lower.

You also can, can see. On some occasions, it's not so frequent, but you can see it also that the milk protein is actually higher than the milk fat, OK? In this case, mostly, ruminal acidosis is, is involved in, in, in, in such cases, you should take the diagnosis a bit further.

For example, you can look at the partition between different particle sizes on the cow's diet by use, for example, of this Penn State particle separator. Or you can assess, or you can assess the ruminal pH, in this case, one of my colleagues, Bruno, is, is the, the guy that works with the nutrition and, and supports management on farm, and he's collecting a ruminal sample with a protected probe, so it's, it's protects it from the saliva contamination with saliva. And it's, he's checking the pH of the of this rumen, of this rumen sam sample to see if the cow is, is, is, has got subacute ruinal acidosis.

OK? So there are different tools to do this as well. And, and normally you, you'll do it if, if there is an indication, in, in terms of milk fat drop in the, in the cows, or in the, on the bulk tank, OK?

OK. So we're, the good news is that we're going on to the final subject, which is diagnosing high total bacteria count. So not necessarily a problem related with mastitis, but it's some, it's an issue that we, every once in a while get called out to check, because The milk, it has a high total bacteria count and the dairy, which is buying, the milk is, is either penalising the or not accepting some, if it comes to that extreme, not accepting the milk anymore, OK?

It normally, it tends to be related with the hygiene of the milk tubes and of the milk tanks. Or also the insufficient or slow cooling of the milk. Mostly, it's one of, one of these, one of these subjects which is involved.

So what we need to check is how fast is the milk, the milk being cooled after it leaves the cow, what kind of system is there, if any, to, to, to drop the, the, the temperature of the milk as soon as it leaves the cow. To check, we want to check also if the temperature is in the bulk tank is maintained constantly around the 4 °C. And also we want to check at what temperature the water that cleans the tank that washes the, the, the tank, and the milking parlour tubes to see if that temperature is sufficiently high, OK?

So, what we tend to use, so sorry, this is, this is, it's Written here in Portuguese, but I've translated it on top. So what we tend to use to assess all these temperatures is is a thermometer with a with a data loger and with a long probe that is, is put into the, into the, into the bulk tank and we leave it on for. Days to see what's what's what's occurring.

So when the truck comes and picks up the milk, what temperature is the water washing the tank? For example, in this situation, you see that the water washing the tank is below 30 degrees, so insufficient. High temperature?

Then, of course, when it's empty, it doesn't really matter what's, what's going on there. Then when the, once the milking starts, how soon does the temperature of the milk drop to around 4 degrees, OK. So we want it to stay, most of the time around 4 degrees, and this is a, this is a good tool to diagnose some of, some of these, some of these issues, OK?

It's, it's fairly It's fairly cheap. You just needed a, a thermometer with a data loger. Leave it on for a few days and then go back and, and, and catch the log and see what's actually going on in this situation, OK?

So we definitely need to check the temperature of the washing solution. We need to check the water quality, so some, very often the water comes from wells, and the quality might or might be different in different times of the year. .

So you need to do bacterial counts in the water as well. You need to check the, the concentrations of the detergents being used against the manufacturer's instructions to see if everything's going according to plan. You can check the slush motion, so the slush is the, the, the turbulence.

Of the water and the air running in inside the tubes, and that could also be measured with with the equipments that measure the vacuum, for example, in the, in the, in the parlour. And also you need to check for areas that might seem a little bit inaccessible for this, for this water. So, so dead ends or curves like, like this, this was actually from a, from a small ruminant milking parlour that had all of this, which is causing, of course, a high total bacteria count in this, in this farm, OK?

What you can actually do also is to sample the, the milk at different times to try to get a better understanding at which point is, is there actually being an increase in the total bacteria count. So, for example, just after the milk, just after leaving the milking parlour, just before it goes into the, into the bulk tank, when it goes out of the bulk tank, so you can sample milk at different time periods, different points rather, and see when is actually the, the, the big increase in, in the total bacteria counts occurring. Certainly, If there is a very high prevalence of mastitis or very, very dirty cows being milk milked, even the best cooling of the best washed.

Properly tank can be overwhelmed. So if you have a very dirty cow or many dirty cows and many cows with mastitis, the total bacteria count may still go up, even if everything is going, everything else is going well. OK?

So in this case, if you sample right after. The milk leaving the cows, you know, into the milk tubes, you'll see that already there is a high total bacteria count, OK? I would recommend, I would recommend if you're interested in these subjects, there's a document online which is called Troubleshooting high bacteria counts in Farm milk, which is issued by the National Mastitis Council, and it, it really is a very, very good guide if you have problems like this.

So, in conclusion, and I'm sorry, I got a little bit over the time that which was allotted to, to me, I would say diagnosing mastitis is, is, is increasingly challenging due to the time labour pressure which is being put on, on farm managers and milkers. Diagnosing mastitis pathogens is, is more widespread, particularly on farm to guide the therapeutic approach. OK.

So, I would recommend using the PCR for initial approach, but also Using, you know, very, widespread, the, the on-farm culture, which is, which allows a very guided therapeutic approach. Looking at the proportion of new infections and, and, and lactating cows or new infections in the dry period or cure rates over the dry period are good measures of the efficacy of preventative measures in place and should be checked monthly or every 3 months, for example, if you're talking about the dry period. The milk recording is a very good snapshot and can give you lots and lots of information all at once, so, please spend some time looking at the milk recording summary if you have it available.

And the last comment I, I would like to, to say in conclusion is that the high total bacteria counts will affect some farms, and most often it's not due to mastitis, but it's more frequently due to poor washing routine or insufficient cooling or insufficient high temperature in the, in, in, in the washing of the parlour or of the bulk tank. Thank you very much for your attention. Thank you to Verbach also for the kind invitation, and I'm open to, to your questions.

Thank you very much. Thank you very much, Ricardo, for your very interesting and detailed presentation on the diagnosis. Yes, we will take some questions now.

Just, I have put on the chat the link to the first episode for those interesting to to watch or watch again the first episode, and we will of course made available the the replay after all. So the question . That was posed, was, what can affect milk density, to, to be lower, especially after it, it's stress.

. Yes, I, I, I would, it's a shame I don't have, Bruno, the, the, my colleague, the nutritionist, here, here with me because, he would probably answer that, straight away. It, it, I, I, I, I wouldn't know how to answer that, with a, with a high degree of certainty.

. It, it, I know that, OK, OK, you need to do, you need to change the, the nutrition of the cow to a higher. More fermentable, less fibrous, diet, normally, . That might change, in itself, it might decrease the fat, but if there is some degree of, of, of, negative energy balance, also you might decrease the protein, but I wouldn't, I wouldn't, I wouldn't risk.

Without, without talking to a nutritionist, some someone that understands a bit more about that than than myself. OK. Other question was what treatment approach you use for a streptoglucose lay in infected form.

OK. Normally it's, it's the only situation in which I would, I recommend treating subclinically affected cows, because that's been proven a long time ago that economically it pays off, to treat cows even with subclinical mastitis, in the herds, for two reasons. 111 being It has a very high cure rate.

So if you treat it with, you know, penicillin or or amoxicillin clavulanic acid, something like that, you will get a cure most often, OK, which is different from Staphhoria, strep ubris, and, and, and the likes of, of pathogens that have lower cure rates. So, fortunately, streptococcus yalactia has a very high very high cure rate. So it works.

The using an antibiotic will, will cure, lead to a cure most often. And because of that, when you're treating one animal, you're actually preventing transmission of infection to other animals. So you're not just affecting that individual animal, you're also affecting other, other animals.

So for that reason it's, it's economically viable to do that. Normally what I would what I would do is I don't recommend treating unless they have sorted their milking routine. So if, unless they're, not.

Force stripping, so I would, I would go into the farm and say, OK, stop force stripping. Use one individual paper towel or cloth to for each cow, . And use some kind of, of liner or cluster spraying with parasitic acid.

Unless they have these trees sorted, I would not go in and recommend treatment because it would just come back, OK? So it's, it, the contagious is still occurring. So once they have those three things sorted and I, and I'm more or less certain that they are not.

Being, contaminated, you know, transmitted between cows, then I would go with treating these cows, and I normally to decrease the The impact on, on, on the farm and of treating many cows at the same time. I normally treat cows in the beginning of lactation up to around 120, because I, those are the ones that are going to be milked for a longer period of time. Those are the ones that are, will give more milk.

So I tend to start treating in the beginning of lactation and leave the ones. Which are in, in the, you know, 2nd and 3rd, thirds of the lactation, I would leave them, because they would be, treated during dry off. I'm not sure if I answer that, but that those would be my comments.

OK. But at the moment there is no additional, questions, so I guess, that will be it for tonight. Thanks again, Ricardo.

Thank you and, and see you in 3 months for the last, episode, that we, we, it will be more about, really the treatment of clinical mastitis. So have a good evening everybody, and thanks again. Bye-bye.

Thank you. Bye-bye.