Scientometrics Evaluation of Published Scientific Papers on the Use of Proteomics Technologies in Mastitis Research in Ruminants.

The objective of this study was the presentation of quantitative characteristics regarding the scientific content and bibliometric details of the relevant publications. In total, 156 papers were considered. Most papers presented original studies (n = 135), and fewer were reviews (n = 21). Most original articles (n = 101) referred to work involving cattle. Most original articles described work related to the diagnosis (n = 72) or pathogenesis (n = 62) of mastitis. Most original articles included field work (n = 75), whilst fewer included experimental (n = 31) or laboratory (n = 30) work. The tissue assessed most frequently in the studies was milk (n = 59). Milk was assessed more frequently in studies on the diagnosis (61.1% of relevant studies) or pathogenesis (30.6%) of the infection, but mammary tissue was assessed more frequently in studies on the treatment (31.0%). In total, 47 pathogens were included in the studies described; most were Gram-positive bacteria (n = 34). The three bacteria most frequently included in the studies were Staphylococcus aureus (n = 55 articles), Escherichia coli (n = 31) and Streptococcus uberis (n = 19). The proteomics technology employed more often in the respective studies was liquid chromatography-tandem mass spectrometry (LC-MS/MS), either on its own (n = 56) or in combination with other technologies (n = 40). The median year of publication of articles involving bioinformatics or LC-MS/MS and bioinformatics was the most recent: 2022. The 156 papers were published in 78 different journals, most frequently in the Journal of Proteomics (n = 16 papers) and the Journal of Dairy Science (n = 12). The median number of cited references in the papers was 48. In the papers, there were 1143 co-authors (mean: 7.3 ± 0.3 co-authors per paper, median: 7, min.-max.: 1-19) and 742 individual authors. Among them, 15 authors had published at least seven papers (max.: 10). Further, there were 218 individual authors who were the first or last authors in the papers. Most papers were submitted for open access (n = 79). The median number of citations received by the 156 papers was 12 (min.-max.: 0-339), and the median yearly number of citations was 2.0 (min.-max.: 0.0-29.5). The h-index of the papers was 33, and the m-index was 2. The increased number of cited references in papers and international collaboration in the respective study were the variables associated with most citations to published papers. This is the first ever scientometrics evaluation of proteomics studies, the results of which highlighted the characteristics of published papers on mastitis and proteomics. The use of proteomics in mastitis research has focused on the elucidation of pathogenesis and diagnosis of the infection; LC-MS/MS has been established as the most frequently used proteomics technology, although the use of bioinformatics has also emerged recently as a useful tool.

Stress related to wild canid predators near dairy sheep farms associated with increased somatic cell counts in bulk-tank milk.

We investigated the association between wild canid predators reported near sheep farms throughout Greece and somatic cell counts in bulk-tank milk as a reflection of milk quality. The study included 325 dairy sheep flocks, where bulk-tank milk somatic cell counts and total bacterial counts were measured and staphylococci were isolated. Farms were divided into three groups: Cohort A (farms with no reports of wild canid predators nearby), B (farms with canid predators (golden jackal and grey wolf) nearby yet with no experience of livestock losses to predation) and C (farms with canid predators nearby and livestock losses to predation). Somatic cell counts in bulk-tank milk of Cohort C farms were significantly higher, + 43% and + 29%, compared to those for Cohorts A and B, respectively: 0.617 × 106 cells mL-1 versus 0.433 × 106 or 0.477 × 106 cells mL-1, respectively. The presence of wild canid predators near sheep farms was associated with lower quality milk potentially indicative of stress consistent with the potential effects of a landscape of fear. Increasing biosecurity measures at livestock farms, e.g., fencing, and presence of livestock guard dogs could minimise predation risk, whilst also improving livestock welfare by reducing predator-associated stress.

Longitudinal Study of Antibiotic Resistance of Staphylococci from Cases of Subclinical Mastitis in Sheep in Greece: Incidence and Risk Factors.

The present paper extends a previous publication on a field study of subclinical mastitis in sheep and focuses on the following laboratory characteristics of the staphylococcal isolates: antibiotic resistance and association with biofilm formation. The specific objectives of the present study were (a) to describe the incidence of isolation of antibiotic-resistant staphylococci from cases of mastitis throughout the milking period in dairy sheep flocks and (b) to identify relevant risk factors, which would contribute to the sustainable control of the infection. Staphylococcal isolates from subclinical mastitis were evaluated for antibiotic resistance to 18 antibiotics. Antibiotic resistance was detected in 57 of the 179 staphylococcal isolates from subclinical mastitis (31.8%). Resistance was recorded against 11 antibiotics, most often against ampicillin (63.2% of resistant isolates), penicillin (63.2%) and tetracycline (47.4%). Isolates resistant to ampicillin and penicillin were recovered in all 12 farms. Twenty-one multidrug-resistant isolates (11.7%) were also recovered. The incidence risk of isolation of staphylococci resistant to at least one (any) antibiotic throughout the study period was 23.8%. The incidence risk of isolation of staphylococci resistant to oxacillin was 5.0%; that of isolation of multidrug-resistant staphylococci was 8.8%. With regard to increased incidence risk of isolation of staphylococci resistant to at least one (any) antibiotic and increased incidence risk of isolation of staphylococci resistant to oxacillin, the omission of anti-staphylococcal mastitis vaccination of ewes emerged as a risk factor. With regard to increased incidence risk of isolation of multidrug-resistant staphylococci, the following variables emerged as risk factors: (a) higher number of antibiotics used on the farm for the treatment of mastitis and (b) younger age of lambs taken away from their dam. Most biofilm-forming antibiotic-resistant staphylococci were recovered from farms where anti-staphylococcal mastitis vaccination was not applied (55.9% versus 44.1% from farms where anti-staphylococcal mastitis vaccination was applied).

Evaluation of the Role of Veterinarians for Outcomes Related to the Health and Production of Dairy Small Ruminants in Greece.

The objective of the present study was to evaluate the potential benefits of veterinarians in improving the health and welfare of dairy sheep and goats by studying the associations of management practices employed in the farms with production- or health-related outcomes in sheep and goat farms in Greece. This work explored associations with 'professional relationship with a veterinarian' at 444 small ruminant dairy farms in an investigation performed around Greece, where 106 variables, related to infrastructure, animals, production outcomes, health management, health problems and human resources, were assessed. In 384 (86.5%) farms, a professional relationship with a veterinarian was maintained. The median value of visits made annually by veterinarians to these farms was five. In farms with a professional relationship with a veterinarian, significant differences were found in 24 variables (35.8%) related to management practices and 6 (30.0%) production- or health-related outcomes. In multivariable analysis, the following emerged with a significant association: epg counts in faecal samples (p = 0.014), average annual milk production per ewe/doe (p = 0.015), somatic cell counts in bulk-tank milk (p = 0.037), and annual incidence of clinical mastitis (p = 0.044). Moreover, associations of the characteristics of veterinarians emerged with somatic cell counts in bulk-tank milk: the gender (p < 0.0001) and the age (p = 0.004) of the veterinarians. The results attest that the application of veterinary advice and clinical services in sheep and goat dairy farms contributes to the improved health, production and welfare of animals.

Irregular Findings on Teatcups in Milking Parlours in Sheep and Goat Farms and Potential Predictors.

The objectives of the present study were as follows: (a) to describe the prevalence of irregular findings on teatcups in milking parlours with dairy sheep and goats after the post-milking cleaning procedures had been completed, (b) to associate staphylococcal isolation from teatcups with the presence with irregular findings and (c) to identify predictors of the presence of irregular findings on teatcups. The teatcups in the milking parlour of 255 sheep and 66 goat farms were macroscopically evaluated for the presence of irregular findings immediately after the completion of cleaning of the parlour. In total, 1115 and 303 teatcups, respectively, were assessed. A detailed interview with the farmer served to record the characteristics of the milking parlour and obtain information about husbandry and health management variables in the farm. Teatcups with macroscopically evident irregular findings were observed in 150 milking parlours (46.7%). Overall, 593 teatcups (41.8%) were found with macroscopically evident irregular findings. Dirt, milk residues and cracks or tears were recorded in the teatcups of 90.0%, 36.0% and 12.7% of parlours with irregular findings. Staphylococci were more frequently isolated from teatcups with irregular findings than from undamaged ones: from 37.4% (222/593) versus 12.8% (106/825). They were more frequently isolated from teatcups with milk residues (39.4%) and teatcups with dirt (39.0%). Via multivariable analysis, the following three variables emerged as significant predictors of presence of teatcups with irregular findings: the daily number of milking sessions, month into the lactation period at sampling and number of available milking units per animal position. The study provides, for the first time internationally, an appraisal of the frequency of problematic teatcups in the milking parlours of small ruminant farms. The analysis of predictors has provided a focus for specific management aspects, where interventions need to be performed, in order to improve the situation in farms with problems. The consequent increased staphylococcal burden on teatcups with irregular findings points to the increased risk of intramammary infections in such cases.

Longitudinal Study of Subclinical Mastitis in Sheep in Greece: An Investigation into Incidence Risk, Associations with Milk Quality and Risk Factors of the Infection.

The objectives of this work were (a) to describe the incidence risk of subclinical mastitis in dairy flocks throughout the milking period, (b) to present potential associations of subclinical mastitis with the quality of milk and (c) to identify risk factors for high-incidence risk of the infection. A longitudinal study was performed in 12 dairy sheep flocks in Greece. Mammary secretion samples from 240 ewes and bulk-tank milk samples were collected in four repeated visits over a period of six months for bacteriological, chemical and cytological examinations. Overall, the incidence risk of subclinical mastitis throughout the study period was 51.7%, and it varied among farms from 25.0% to 75.0%. The respective figure for staphylococcal subclinical mastitis was 48.8%. The incidence risk of recurrence of subclinical mastitis among ewes in the flocks was 35.4%. The most frequently identified bacteria from cases of subclinical mastitis were S. aureus and S. simulans; of the mastitis-causing staphylococcal isolates, 65.4% were biofilm-forming. Somatic cell counts in bulk-tank milk progressively increased as the lactation period advanced, with significant increases seen on the third and fourth visits to the farms. Somatic cell counts in bulk-tank milk correlated well with prevalence of subclinical mastitis in flocks. A significant inverse correlation and a significant positive correlation were seen for prevalence of subclinical mastitis versus total protein content or added water in bulk-tank milk. During multivariable analysis, younger age of newborns when taken away from the dam and consequently delayed start of milking of ewes, omission of anti-mastitis vaccination of ewes and lack of employed staff on the farms emerged to be significantly associated (positively) with increased incidence risk of subclinical mastitis.

Recovery of Staphylococci from Teatcups in Milking Parlours in Goat Herds in Greece: Prevalence, Identification, Biofilm Formation, Patterns of Antibiotic Susceptibility, Predictors for Isolation.

The objectives of this work are (a) to describe staphylococci on the teatcups of milking parlours in goat farms and identify predictors for the presence of staphylococcal isolates on the teatcups, (b) to evaluate relationships with total bacterial counts and somatic cell counts in bulk-tank milk, and (c) to establish patterns of susceptibility to antibiotics for the staphylococcal isolates and identify predictors for the recovery of resistant isolates. In a cross-sectional study of 66 goat farms across Greece, swab samples were collected from 303 teatcups (upper and lower part) for staphylococcal recovery, identification, and assessment of biofilm formation. Details regarding health management on the farms (including conditions in the milking parlour) and the socio-demographic characteristics of farmers were collected by means of a structured questionnaire. A total of 87 contaminated teatcups (28.7%) were found on 35 goat farms (53.0%). Staphylococci were more frequently recovered from the upper than the lower part of teatcups: 73 versus 43 teatcups, respectively. After identification, 67 staphylococcal isolates (i.e., excluding similar isolates) were recovered from the teatcups; Staphylococcus aureus, Staphylococcus capitis, and Staphylococcus equorum predominated. Of these isolates, 82.1% were biofilm-forming. In multivariable analysis, the annual incidence of clinical mastitis in the herd emerged as the only significant factor associated with the isolation of staphylococci from the teatcups. Of the 67 isolates, 23 (34.3%) were resistant to at least one antibiotic, and 14 (22.4%) were multi-resistant. Resistance was found most commonly against penicillin and ampicillin (22.4% of isolates), fosfomycin (17.9%), clindamycin (14.9%), erythromycin, and tetracycline (13.4%). In multivariable analysis, the annual incidence of clinical mastitis in the herd and the use of detergent for parlour cleaning emerged as significant factors associated with the isolation of staphylococci resistant to antibiotics.

Isolation of Biofilm-Forming Staphylococci from the Bulk-Tank Milk of Small Ruminant Farms in Greece.

The objectives of this study were (i) to describe staphylococcal isolates recovered from bulk-tank raw milk collected from sheep and goat farms during a countrywide study performed in Greece, (ii) to study management factors potentially associated with their presence in bulk-tank milk and (iii) to provide evidence regarding their association with the quality of the milk. In total, 312 staphylococcal isolates, recovered from samples of bulk-tank raw milk from 444 small ruminant farms in Greece, were evaluated in this work. The in vitro formation of biofilm by the isolates was tested by combining the findings of (a) culture appearance on Congo Red agar plates and (b) results of a microplate adhesion test. The most frequently identified species was Staphylococcus aureus (75 isolates); other frequently recovered species were S. simulans (44 isolates), S. equorum (34 isolates) and S. haemolyticus (26 isolates); in total, 23 species were identified. In total, 224 (71.8%) isolates were biofilm-forming and were recovered from the bulk-tank milk samples of 148 sheep flocks (45.5%) and 55 goat herds (46.2%). There was evidence of seasonality in the isolation of staphylococci: during spring, mostly biofilm-forming isolates were recovered, whilst during summer, mostly non-biofilm-forming isolates were recovered. Among farms applying machine-milking, the proportion of farms from which biofilm-forming isolates were recovered was higher where water with temperature < 50 °C or ≥90 °C was used to clean the milking parlour. In the multivariable analyses, for farms applying machine-milking, the temperature of the water emerged as the only significant variable (p = 0.024), whilst in farms applying hand-milking, the only tendency that emerged was for the frequency of collection of milk from the farm tank (p = 0.08). In sheep flocks, recovery of biofilm-forming staphylococci from the bulk-tank milk was associated with higher somatic cell counts and higher total bacterial counts in the milk. The study identified abiotic factors related to the presence and isolation of these bacteria, specifically the temperature of water used for the cleaning of the milking parlour (in farms where machine-milking is applied) and the frequency of milk collection from the farm tank. These factors apply after the production of milk, and they could thus be regulated appropriately in order to reduce bacterial load and improve the quality of milk delivered to dairy plants. In sheep farms, an association was also seen between recovery of biofilm-forming staphylococci and high somatic cell counts in milk.

Data on Mapping 444 Dairy Small Ruminant Farms during a Countrywide Investigation Performed in Greece.

The small ruminant industry is the most important branch of Greek agriculture. Nevertheless, despite the importance of small ruminant farming for Greece, no detailed mapping of the respective farms has been undertaken and published. The present work refers to mapping the dairy sheep and goat industry in Greece through an extensive, countrywide cross-sectional study, in which information was obtained from 325 sheep and 119 goat farms. The objectives were the collection, the classification and the presentation of data obtained from all these farms through interviews, using a questionnaire and through examination of samples collected during the visits. All the farms enrolled in the study were visited by the investigators. Initially, information was obtained by means of a detailed, structured questionnaire with 442 questions. Moreover, samples of milk were collected from the bulk-tank of each farm and faecal samples were collected from female animals in each farm. The milk samples were processed for cytological and microbiological examination. Staphylococcus spp. and Listeria spp. isolates were recovered and identified at a species level; furthermore, a full antibiotic sensitivity pattern assessment was conducted. Faecal samples were processed by standard parasitological tests for the identification of protozoan, trematode, cestode and gastrointestinal and respiratory nematode parasites. The paper presents the cumulative findings of the study, i.e., the answers to the questions during the interviews and the results of the laboratory examinations performed in the samples; the findings are presented separately for sheep and goat farms. The findings can be used as baseline measurements; individual farms and cohorts of farms can be compared against the current results to draw conclusions against the countrywide situation. Moreover, the current results can be used as reference points for the future, in order to assess changes that might have occurred in the meantime. The study also described the differences between farms with sheep or goats; in total, differences in 137 parameters were identified. Some of these can be attributed to the different management systems practiced; sheep flocks are managed mostly under the intensive or semi-intensive system, whilst goat herds are managed mostly under the semi-extensive or extensive system. These findings can be useful in the health management of small ruminants, in providing evidence-based support and within the scope of precise livestock medicine and health management.

Helminth Infections in Dairy Sheep Found in an Extensive Countrywide Study in Greece and Potential Predictors for Their Presence in Faecal Samples.

The aims of the present study were: (a) to describe the prevalence of helminth infections from pooled faecal samples from sheep flocks across Greece and (b) to evaluate flock-related factors potentially associated with the presence of these infections in the flocks. An extensive countrywide study was performed on 325 sheep farms throughout Greece; faecal samples were collected from ewes and processed for the identification of helminth parasites. Helminths were detected in samples from 92.9% of flocks; these included Dicrocoelium dendriticum (16.7% of flocks), Fasciola hepatica (0.6%), Paramphistomum cervi (2.2%), Moniezia spp. (18.8%), Trichostrongylidae (85.5%), Nematodirus spp. (18.8%), Strongyloides papillosus (7.1%), Trichuris spp. (20.0%) and lungworms (17.8%). Mean Trichostrongylidae counts across all flocks in the study were 215 epg. Specifically, for Trichostrongylidae, there were differences between flocks that had or had not received anthelmintics during the two months prior to sampling, as well as between flocks located in various areas of the country. In multivariable analyses, for the outcome 'high (>300) epg counts in faecal samples', the month into the lactation period at sampling and the application of reproductive control practices on the farm emerged as significant factors; for the outcome 'high proportion (>63%) of Teladorsagia spp. in faecal samples', the availability of straw bedding emerged as a significant factor; and for the outcome 'high proportion (>63%) of Haemonchus contortus in faecal samples', the age of the farmer emerged as a significant factor. For the outcome 'presence of Trichuris spp. in faecal samples', the provision of finished feed (concentrate) to animals emerged as a significant factor, whilst, finally, for the outcomes 'presence of D. dendriticum in faecal samples' and 'presence of lungworms in faecal samples,' no significant factors emerged.

Patterns of Reproductive Management in Sheep and Goat Farms in Greece.

This paper reports findings regarding patterns of reproductive management in 325 sheep flocks and 119 goat herds throughout Greece. The objectives were to describe the patterns of reproductive management in sheep and goat farms, to highlight factors that were associated with these management patterns and the clinical application of the various reproductive management approaches in the farms and to establish potential associations with production outcomes. The median months of the start of the mating period for adult sheep and goats were May and June, respectively and for ewe lambs and replacement goats these were August and September, respectively. The median duration of the mating period was 2 months for adults and 1 month for replacements. The median ratio of female to male animals was 22:1 and 25:1 in sheep and goat farms, respectively. Control of reproduction was applied in 33.2% and 16.8% of sheep and goat farms, respectively, mostly by intravaginal progestagen sponges and less often by melatonin implants; multivariable analysis indicated five (availability of milking parlour, number of ewes in a farm, number of daily milkings, daily period spent by farmers at the farm, farming tradition in the family) and two (number of daily milkings, availability of milking parlour) predictors in sheep and goat farms, respectively. Pregnancy diagnosis ultrasonographically was performed in 36.6% and 16.8% of sheep and goat farms, respectively; multivariable analysis indicated as significant three (management system applied in farms, age of farmer, farming tradition in the family) and two (management system applied in farms, availability of milking parlour) predictors in sheep and goat farms, respectively. The control of reproduction and pregnancy diagnosis were associated with a higher number of vaccinations during pregnancy: 2.6 and 2.7, respectively, versus 2.2 and 2.1 in farms where they were not performed. The average per farm number of lambs/kids born per female animal was 1.33 ± 0.01 and 1.30 ± 0.02 for sheep and goat farms, respectively; five (control of reproduction, location of the farm, presence of working staff, collaboration with veterinary practice, breed of ewes) and two (control of reproduction, breed of female goats) predictors were identified in sheep and goat farms, respectively, for high numbers of lambs/kids. Lambs/kids were taken away from their dams at the age of 50 and 65 days, respectively. The average culling age (females, males) was 5.9 and 4.4 years and 6.9 and 4.9 years for sheep and goats, respectively. Farmers sourced animals to be used as replacements for those removed from the farm, mostly from the animals in their own farms, considering criteria related to milk production. Finally, the application of the control of reproduction was associated with increased milk production and the number of newborns, whilst an inverse correlation between the culling age of animals and milk production was seen.

Vaccination against Bacterial Mastitis in Sheep.

The objective of this review is to discuss the application of vaccination for the prevention of bacterial mastitis in ewes, performed within the frame of health management schemes in sheep flocks. Mastitis is a multi-faceted infection, caused most often by staphylococci; hence, special emphasis is given to staphylococcal mastitis, also given that most relevant studies refer to vaccinations against that infection. Studies regarding various vaccines have been performed; most studies refer to vaccination by using a vaccine making use of cell-free surface polysaccharides in various vehicles, bacterial unbound cells or bacterial cells embedded in their biofilm matrix. Vaccination against mastitis should be better performed during the final stage of pregnancy to allow protection of ewes from lambing and should be considered as one of many control measures for the prevention of the disease. The expected benefits of mastitis vaccination in sheep flocks include the following: (a) reduced incidence risk of clinical and subclinical mastitis, (b) reduced somatic cell counts, optimum chemical composition, absence of staphylococci in milk, (c) increased milk production, (d) reduced dissemination of mastitis-causing pathogens and (e) reduction of antibiotic use in flocks.

Scientometrics Study of Research Output on Sheep and Goats from Greece.

The study is a scientometrics evaluation of published articles performed in Greece on sheep and goats during the last 25 years, a period coinciding with implementation of reforms to shape and consolidate tertiary education and research establishments in the country. Objectives were: evaluation of the relevant publications and presentation of quantitative characteristics regarding scientific content and bibliometric details. The Web of Science platform was used (search terms: [[sheep OR goat*] OR [Ovis aries OR Capra hircus]] (1997-2022)) and 1080 papers were considered in detail. Throughout the study period, there was a clear progressive increase in numbers of papers published. The papers originated from 39 different entities, most from Aristotle University of Thessaloniki (34.0%), University of Thessaly (28.0%), Agricultural University of Athens (21.2%) and Hellenic Agricultural Organization-Dimitra (13.6%). Papers were published in 318 different journals. Journals with more published papers were Small Ruminant Research and Journal of the Hellenic Veterinary Medical Society. The most frequent general topic of study in the papers was health and welfare (46.7% of papers); second most frequent topic was animal products (18.6%). The papers have received 16,558 citations, i.e., οn average 15.4 citations per paper; the h-index was 56, the i10-index was 518 and the yearly citations per paper were 1.71. Papers on goats had higher impact than papers on sheep. There were 1711 individual authors, of which 728 were first or last authors. In total, 24 authors have each co-authored ≥2.5% of all papers; five authors were each first or last in that proportion of all papers. The findings have indicated that relevant research has focused on milk production and diseases of the udder of small ruminants; moreover, there was accumulation of relevant research in Greece in some establishments only. The findings of the study can be employed to initiate improved relevant research approaches in the country.

Protozoan Parasites in Adult Dairy Small Ruminants and Potential Predictors for Their Presence in Faecal Samples.

There is a scope to study protozoan infections in adult ewes and does, as these animals can act as reservoirs of infection for lambs and kids, for which these pathogens are harmful. The objectives of this study were to describe the prevalence of protozoan infections in faecal samples from adult sheep and goats on dairy farms across Greece and to evaluate farm-related factors potentially associated with the presence of protozoan infections in these animals. A cross-sectional study was performed on 325 sheep and 119 goat farms throughout Greece; faecal samples were collected from ewes and does and processed for the identification of protozoan parasites. Eimeria oocysts were found in faecal samples from 69% of farms (72% of sheep farms and 61% of goat farms), Giardia cysts in samples from 33% of farms (33% of sheep farms and 34% of goat farms) and Cryptosporidium oocysts in samples from 8% of farms (7% of sheep farms and 11% of goat farms). In a multivariable analysis, for the presence of Eimeria in samples from sheep farms, the lack of a designated building for lambs emerged as a significant factor; for the presence of Giardia in samples from goat farms, the availability of a main building for animals emerged as a significant factor; for the presence of Cryptosporidium, the lack of grazing and the management system emerged as the main significant factors in sheep and goat farms, respectively. Protozoa were found significantly more frequently in samples collected from farms on which farmers considered diarrhoea as an important health problem in their lambs/kids.

Administration of Vaccines in Dairy Sheep and Goat Farms: Patterns of Vaccination, Associations with Health and Production Parameters, Predictors.

This paper reports findings regarding patterns of vaccine usage in sheep and goat farms, in 325 sheep flocks and 119 goat herds throughout Greece. The objectives of the study were (a) to describe the patterns of vaccine administration in small ruminant farms and (b) to highlight factors that were associated with vaccinations in the farms. Vaccination against brucellosis was performed in all farms into the study. Among optional vaccinations, anti-clostridial vaccination was most frequently performed (in 97.8% of farms), followed by vaccination against contagious agalactia, (56.5% of farms), pneumonia (41.2%), chlamydial abortion (38.1%), staphylococcal mastitis (36.0%), and paratuberculosis (9.5%). Vaccinations against pneumonia and staphylococcal mastitis were performed more frequently in sheep flocks, whilst vaccinations against paratuberculosis were performed more frequently in goat herds. On average, 2.8 and 2.7 optional vaccinations (i.e., additionally to vaccination against brucellosis) were performed in sheep and goat farms, respectively. The increased number of vaccines administered was associated with a higher average milk production in the respective farms. There was an association of vaccination against staphylococcal mastitis with a reduced recovery of staphylococci from the bulk-tank raw milk. In multivariable analyses, significant associations of the administration of the various optional vaccines were seen with 15 variables, 11 related to health management practices and 4 related to the demographic characteristics of farmers; the collaboration with a veterinarian, the daily number of milking sessions, and the period spent daily by the farmer at the farm premises were each associated with the administration of vaccines against three infections.

Zoonotic Problems Reported by Sheep and Goat Farmers and Factors Potentially Contributing to the Occurrence of Brucellosis among Them.

The study aimed to investigate the occurrence of zoonotic problems reported by dairy small ruminant farmers in Greece and to study potential associations with socio-demographic characteristics of the farmers and management practices applied in the farms. A countrywide investigation was performed in 325 sheep and 119 goat farms in the 13 administrative regions of Greece. The selected farms were visited and interviews were conducted with respective farmers. The occurrence of zoonotic problems in the farmers was recorded. A total biosecurity score (0−6) was devised, based on biosecurity practices followed in farms. Sixty-seven farmers (15.10%, 95% confidence intervals (CI): 12.1−18.7%) reported experiencing a zoonotic problem. Most of the farmers (n = 57) (85.1%, 95% CI: 74.76−91.7%, of those with a zoonotic problem) (12.8%, 95% CI: 10.0−16.3%, of all) reported that the zoonotic problem had been brucellosis. Odds ratio for the occurrence of brucellosis in goat farmers was 1.879 (95% CI: 1.051−3.359) compared to the occurrence of the infection in sheep farmers (p = 0.033). For the outcome 'occurrence of brucellosis' in sheep farmers, the application of hand-milking, the availability of a separate lambing area and the presence of cats in the farm emerged as significant (p < 0.01); for the same outcome in goat farmers, only the availability of a separate kidding area emerged as significant (p = 0.001). The mean biosecurity score in farms in the continental area of the country was significantly higher than in the islands: 3.45 ± 0.05 versus 2.76 ± 0.28, respectively (p = 0.006), whilst there was also a significantly higher score in farms, where the farmer reported occurrence of brucellosis: 3.68 ± 0.15 versus 3.34 ± 0.06 in farms, where the farmer did not report such an incident (p = 0.042). In farms, where the above predictors prevail, farmers should be warned of an increased potential risk for human infection and biosecurity measures should be implemented and tightened.

Isolation of Listeria ivanovii from Bulk-Tank Milk of Sheep and Goat Farms-From Clinical Work to Bioinformatics Studies: Prevalence, Association with Milk Quality, Antibiotic Susceptibility, Predictors, Whole Genome Sequence and Phylogenetic Relationships.

A cross-sectional study was performed in 325 sheep and 119 goat dairy farms in Greece. Samples of bulk-tank milk were examined by standard microbiological techniques for Listeria spp. Listeria monocytogenes was isolated from one (0.3%) and Listeria ivanovii from three (0.9%) sheep farms. No associations between the isolation of L. monocytogenes or L. ivanovii and milk quality were found. No resistance to antibiotics was identified. Three variables emerged as significant predictors of isolation of the organism: the presence of pigs, low average relative humidity and a high number of ewes on the farm. The three L. ivanovii isolates were assessed in silico for identification of plasmids, prophages, antibiotic resistance genes, virulence factors, CRISPRs and CAS genes. Phylogenetic analysis using the core genome revealed that the three strains belonged to the L. ivanovii subsp. ivanovii branch and were especially close to the PAM 55 strain. All strains of the branch appeared to be very similar, with the distance between them being small.

Use of Antibiotics against Bacterial Infections on Dairy Sheep and Goat Farms: Patterns of Usage and Associations with Health Management and Human Resources.

The objectives of the study were (a) to describe the patterns of antibiotic usage against four major clinical problems and (b) to evaluate factors that were associated with their use on small ruminant farms. Sheep and goat farmers mostly administered the antibiotics to animals at the dose prescribed (80.4%) and observed the necessary withdrawal period (98.7%), but fewer farmers (22.3%) weighed the animals to calculate their bodyweight before antibiotic administration. For the treatment of clinical mastitis, oxytetracycline, penicillin and streptomycin were the antibiotics used more frequently; 2.03 different antibiotics were used per sheep flock and 2.06 per goat herd, most frequently administered in injectable forms (88.8% of farms). In cases of abortion, oxytetracycline was administered more frequently; 1.12 different antibiotics were used per sheep flock and 1.03 per goat herd. In 94 farms (21.2%), routine administration of antibiotics was performed to newborns; oxytetracycline and ampicillin were administered more often. For the treatment of pneumonia in newborns, oxytetracycline, penicillin and tulathromycin were used more frequently; 1.33 antibiotics were used per sheep flock and 1.29 per goat herd. For the treatment of diarrhoea in lambs and kids, oxytetracycline, amoxicillin and penicillin were the antibiotics used more frequently; 1.34 antibiotics were used per sheep flock and 1.59 per goat herd. Results of multivariable analyses indicated 16 variables associated with the various outcomes for usage of antibiotics for the treatment of the above clinical problems. Of these, 11 variables were associated with the farmers: education of farmers was significant for three outcomes; the age, the experience, the professional involvement and farming family tradition of farmers and the daily period spent at the farm were each significant for one outcome.

Detection of Zoonotic Gastrointestinal Pathogens in Dairy Sheep and Goats by Using FilmArray® Multiplex-PCR Technology.

The objectives of this study were (a) to detect gastrointestinal pathogens in faecal samples of sheep and goats using the FilmArray® GI Panel and (b) to evaluate factors that were associated with their presence. Faecal samples from ewes or does in 70 sheep flocks and 24 goat herds in Greece were tested for the presence of 22 gastrointestinal pathogens by means of the BioFire® FilmArray® Gastrointestinal (GI) Panel. The most frequently detected pathogens were Shiga-like toxin-producing Escherichia coli stx1/stx2 (94.7% of farms), Giardia lamblia (59.6%), and Campylobacter spp. (50.0% of farms). Other pathogens detected were Cryptosporidium spp., Salmonella spp., enterotoxigenic E. coli lt/st, Yersinia enterocolitica, E. coli O157, Rotavirus A, Shigella/enteroinvasive E. coli, and Plesiomonasshigelloides. There was a difference in the prevalence of detection of pathogens between sheep and goat farms only for Salmonella spp.: 18.3% versus 0.0%, respectively. Mixed infections were detected in 76 farms (80.9% of farms), specifically 57 sheep flocks and 19 goat herds, with on average, 2.5 ± 0.1 pathogens detected per farm. The body condition score of ewes in farms, in which only one pathogen was detected in faecal samples, was significantly higher than that of ewes in farms, in which at least two pathogens were detected: 2.55 ± 0.11 versus 2.31 ± 0.04. In sheep flocks, the number of pathogens in faecal samples was significantly higher in farms with semi-extensive management. In goat herds, the number of pathogens in faecal samples was positively correlated with average precipitation and inversely correlated with temperature range in the respective locations.

High Milk Somatic Cell Counts and Increased Teladorsagia Burdens Overshadow Non-Infection-Related Factors as Predictors of Fat and Protein Content of Bulk-Tank Raw Milk in Sheep and Goat Farms.

This paper presents the results of an extensive countrywide investigation performed in 325 dairy sheep flocks and 119 goat herds throughout Greece. The objectives of the study were (a) to investigate fat and protein content in the bulk-tank raw milk of small ruminant farms in Greece and (b) to identify factors potentially influencing that content and factors that can contribute to increased content. The mean fat/protein contents in bulk-tank raw milk of sheep and goats were 6.16 ± 0.05%/4.43 ± 0.01% and 4.77 ± 0.44%/3.23 ± 0.30%, respectively. Significant differences were seen in protein content between farms in the various parts of the country. For sheep, multivariable analyses revealed breed and age of lamb removal from dams as significant for fat content, and somatic cell counts, management system in the farm, administration of anthelmintic treatment during the last stage of pregnancy, and farmer education as significant for protein content. For goats, significant factors were month into lactation period, age of kid removal from dams, and breed for fat content, and somatic cell counts, month into lactation, grazing duration, and % Teladorsagia larvae in faecal samples for protein content. For concurrently high fat and protein content, in multivariable analyses, the following emerged as significant factors: somatic cell counts in milk, numbers of parasite eggs in faeces, and veterinary collaboration (sheep), and month into lactation and somatic cell counts in milk (goats). The results indicate that high somatic cell counts in milk (reflecting the presence of mastitis) and gastrointestinal parasitic infections (mainly Teladorsagia infection) appear to exert a more significant influence on fat and protein content of milk, in comparison to non-infection-related factors.