Seroprevalence and herd-level associated factors of pathogenic Leptospira spp. circulating locally in dairy cattle in Uruguay.

Leptospirosis is a zoonotic disease of worldwide importance. In Uruguay, it is endemic in cattle and primarily affects people with occupational exposure to livestock. The aim of this study was to determine the national seroprevalence and associated factors of local pathogen Leptospires in dairy cattle. A cross-sectional study was carried out. Herds were stratified by size (1-50, 51-250, and > 250 cattle), and up to 60 dairy cows per herd were randomly selected. A total of 4269 serum samples from 101 dairy herds were analyzed by microscopic agglutination test (MAT). A two-stage sampling design was used to estimate population seroprevalence of Leptospira spp. In order to determine the factors associated with the disease, herds with at least 1 seropositive animal were considered as case herds. Seroprevalence of Leptospira was 27.80% with a 95% CI [21.06, 34.54] at the animal level and 86.92% with a 95% CI [80.00, 93.75] at the herd level. The serology confirms the predominance of serogroups Sejroe and Pomona in our herd with the presence of incidental leptospires infection, in smaller proportion, but with a wide distribution at farm level. The population size and purchasing replacement of cows on dairy farms were associated with infection at farm level. The serologic studies confirmed that exposure to Leptospira spp. is endemic in our herds, and the spreading over dairy herds. Although the movement of purchased females and the size of the herd were associated with the disease, more studies should be conducted, to better understand the epidemiology of the disease and to highlight the possible risks to public health, especially in rural workers, farmers and veterinarians.

Serovars, Virulence and Antimicrobial Resistance Genes of Non-Typhoidal Salmonella Strains from Dairy Systems in Mexico.

Salmonella isolated from dairy farms has a significant effect on animal health and productivity. Different serogroups of Salmonella affect both human and bovine cattle causing illness in both reservoirs. Dairy cows and calves can be silent Salmonella shedders, increasing the possibility of dispensing Salmonella within the farm. The aim of this study was to determine the genomic characteristics of Salmonella isolates from dairy farms and to detect the presence of virulence and antimicrobial resistance genes. A total of 377 samples were collected in a cross-sectional study from calves, periparturient cow feces, and maternity beds in 55 dairy farms from the states of Aguascalientes, Baja California, Chihuahua, Coahuila, Durango, Mexico, Guanajuato, Hidalgo, Jalisco, Queretaro, San Luis Potosi, Tlaxcala, and Zacatecas. Twenty Salmonella isolates were selected as representative strains for whole genome sequencing. The serological classification of the strains was able to assign groups to only 12 isolates, but with only 5 of those being consistent with the genomic serotyping. The most prevalent serovar was Salmonella Montevideo followed by Salmonella Meleagridis. All isolates presented the chromosomal aac(6')-Iaa gene that confers resistance to aminoglycosides. The antibiotic resistance genes qnrB19, qnrA1, sul2, aph(6)-Id, aph(3)-ld, dfrA1, tetA, tetC, flor2, sul1_15, mph(A), aadA2, blaCARB, and qacE were identified. Ten pathogenicity islands were identified, and the most prevalent plasmid was Col(pHAD28). The main source of Salmonella enterica is the maternity areas, where periparturient shedders are contaminants and perpetuate the pathogen within the dairy in manure, sand, and concrete surfaces. This study demonstrated the necessity of implementing One Health control actions to diminish the prevalence of antimicrobial resistant and virulent pathogens including Salmonella.

Environmental and Breed Risk Factors Associated with the Prevalence of Subclinical Mastitis in Dual-Purpose Livestock Systems in the Arauca Floodplain Savannah, Colombian Orinoquia.

The aim of this study was to assess the environmental and breed risk factors associated with the prevalence of subclinical mastitis (SCM) in cows in the dual-purpose livestock system of Arauca, Colombian Orinoquia. Milk samples were taken from 1924 mammary quarters, corresponding to 481 cows on 28 different farms, and the California Mastitis Test (CMT) was applied. Risk factors associated with SCM were determined using multiple logistic regression analysis. The response variable was the presence (1) or absence (0) of SCM. Breed was included as a genetic risk factor, and daily milk production, number of cows in production, lactation month, calving number, cow age, climatic period, and body condition were included as environmental risk factors. The analysis of the odds ratio (OR) of significant effects indicated that the factors significantly associated with the presence of SCM were the number of cows (OR = 2.29; p = 0.005), milk production (OR = 0.88; p = 0.045), and the Taurus-Indicus breeds (OR = 1.79; p = 0.009) and composite breed (OR = 3.95; p = 0.005). In this study, the occurrence of SCM was determined by the following risk factors: number of cows, milk production, and breed. Likewise, the highest prevalence seemed to occur on farms with less technological development and sanitary management of producers from the lowest socioeconomic stratum.

Exploring Antimicrobial Resistance Profiles of E. coli Isolates in Dairy Cattle: A Baseline Study across Dairy Farms with Varied Husbandry Practices in Puerto Rico.

Antimicrobial treatment in livestock can contribute to the emergence and spread of antimicrobial-resistant (AMR) microorganisms. Despite substantial surveillance of AMR bacteria in the continental United States, the prevalence of these AMR organisms in U.S. territories, such as Puerto Rico, remains understudied. The goals of this research included obtaining baseline data on the antimicrobial profile of E. coli isolates from Puerto Rico dairy farms with different husbandry practices. Seventy-nine fecal samples were collected from two types of conventional dairy farms: those that fed calves with tank milk and those that fed calves with waste milk. These samples were collected from the animals' rectums, culture, and subsequently confirmed through biochemical tests. Out of these samples, 32 isolates were analyzed phenotypically and genotypically to elucidate their AMR profiles. The results underscore a discrepancy in the occurrence of antimicrobial resistance genes between calves and adult cattle. Notably, waste milk-fed calves exhibited a significantly higher prevalence of antibiotic-resistant E. coli when compared to their tank milk-fed counterparts. These disparities emphasize the need for more comprehensive investigations to determine causative factors. These results underscore the urgency of comprehensive strategies to raise awareness about how management practices influence antimicrobial resistance, shifting the focus from treatment to prevention.

Groundwater quality and vulnerability in farms from agricultural-dairy basin of the Argentine Pampas.

Agricultural and livestock activities strongly influence groundwater quality and conditioning its use as water supply in rural areas. The aim was to determine the quality and suitability of the groundwater supply used in dairy farms of an agricultural area of Pampa plain of Córdoba (Argentina). Piper's diagram showed that the groundwater types were sodium bicarbonate, sodium bicarbonate-chloride, sodium chloride-sulfate, and sodium sulfate. Physicochemical parameters revealed that cations and anions showed a high and significant correlation in water samples, indicating a strong water-rock interaction. Nitrate (NO3-) content was significantly correlated with pH, water well depth, and distance from contamination sources. A high positive correlation between arsenic (As) and bicarbonate, sulfate, sodium, and chloride (p < 0.05) indicates a similar origin. Among the pesticides monitored, 2,4-D was detected in 25% of groundwater samples (0.4 to 0.8 µg/L) coinciding with the ordinary application practices. In general, most of the groundwater samples did not comply with national and international regulations for drinking water and dairy hygiene, due to the high content of As, NO3-, bacteria, and the presence of 2,4-D herbicide. However, the quality of water was suitable for livestock drink. The data obtained in this study contribute to a better understanding of the contamination processes taking place and improve the agricultural and livestock management for an efficient use of this resource by dairy farmers.

Milk production process, quality and technological properties of milk for the use of various types of milking machines

Studies have been carried out to evaluate the efficiency of using easy-assembled cow houses in modern milk production technologies for the use of high-productive milking plants of the type ‘Parallel and ‘Carousel with 32 machines each. It has been established that new types of premises not only provide comfortable conditions for the maintenance of highly productive cows, but also reduce the labor costs for their maintenance and, most importantly, allow the use of modern high-productive milking installations of the type ‘Parallel and ‘Carousel. It has been established that the technology of preparation of cows for milking and milking technology provides more complete display of the milk ejection reflex at the installation of the ‘Parallel type. The average intensity of cows milk flows at this plant is 30% higher compared to those of the installation of ‘Carousel type, which is confirmed by the intensity of milk flow production at the first minute of milking, which is at the level of 2.97 against 1.85 kg min.-1 per installation of ‘Carousel type. Milk obtained using a milking installation of the ‘Parallel type has higher values of the mass fraction of fat and protein that is associated with the genetic potential of animals. According to physical, chemical and technological properties, milk obtained from milking installations such as ‘Parallel and ‘Carousel is within the limits of the standards in force. Milk obtained from the use of milking equipment such as ‘Carousel has higher electrical conductivity at the level of 4.6 mS cm-3, which is confirmed by a higher level of mastitis disease of cows. Due to bacterial contamination, reductase test and milk clot characteristic, milk obtained using a milk installation of ‘Parallel type also has higher quality indicators than the installation of ‘Carousel type. But according to the complex of indicators, milk obtained from various technologies of milking refers to the desired cheese-making class.(AU)

Milk production process, quality and technological properties of milk for the use of various types of milking machines

Studies have been carried out to evaluate the efficiency of using easy-assembled cow houses in modern milk production technologies for the use of high-productive milking plants of the type ‘Parallel’ and ‘Carousel’ with 32 machines each. It has been established that new types of premises not only provide comfortable conditions for the maintenance of highly productive cows, but also reduce the labor costs for their maintenance and, most importantly, allow the use of modern high-productive milking installations of the type ‘Parallel’ and ‘Carousel’. It has been established that the technology of preparation of cows for milking and milking technology provides more complete display of the milk ejection reflex at the installation of the ‘Parallel’ type. The average intensity of cows’ milk flows at this plant is 30% higher compared to those of the installation of ‘Carousel’ type, which is confirmed by the intensity of milk flow production at the first minute of milking, which is at the level of 2.97 against 1.85 kg min.-1 per installation of ‘Carousel’ type. Milk obtained using a milking installation of the ‘Parallel’ type has higher values of the mass fraction of fat and protein that is associated with the genetic potential of animals. According to physical, chemical and technological properties, milk obtained from milking installations such as ‘Parallel’ and ‘Carousel’ is within the limits of the standards in force. Milk obtained from the use of milking equipment such as ‘Carousel’ has higher electrical conductivity at the level of 4.6 mS cm-3, which is confirmed by a higher level of mastitis disease of cows. Due to bacterial contamination, reductase test and milk clot characteristic, milk obtained using a milk installation of ‘Parallel’ type also has higher quality indicators than the installation of ‘Carousel’ type. But according to the complex of indicators, milk obtained from various technologies of milking refers to the desired cheese-making class.

Epidemiological study of neosporosis in Uruguayan dairy herds.

Neosporosis is one of the leading causes of abortion in cattle worldwide, posing a great economic burden on cattle producers. The aim of this study was to determine the national seroprevalence and putative risk factors of Neospora caninum (N. caninum) in dairy cattle in Uruguay. A cross-sectional study was carried out. Herds were stratified by size (1-50, 51-250, and >250 cattle) and up to 60 dairy cows per herd were randomly selected. Four thousand two hundred twenty-three serum samples from 102 dairy herds were analyzed by indirect ELISA test, under the manufacturer's recommendations. In addition, the herdsman was surveyed and a population study was carried out. The in degree data, geographical coordinates, and seroprevalence of bovine viral diarrhea, enzootic bovine leukemia and infectious bovine rhinotracheitis were available for each herd. A sampling design was used to estimate population seroprevalence of N. caninum. In order to determine the factors associated with the disease, herds with an intra-herd seroprevalence over 20% were considered as case herds. Seroprevalence of N. caninum was 22.3% (95% CI: 18.7-25.9 %) and 96.0% (95% CI: 92.1-99.8%) at the animal and herd level, respectively. The number of dogs on the dairy farms were associated with infection levels (OR: 1.43, 95% CI: 1.02 to 2.03). It was concluded that N. caninum is endemic in the country, and is spreading over dairy herds. Although this study showed evidence that the number of dogs were associated with high levels of infection, more studies should be conducted, to better understand the epidemiology of the disease and thus develop efficient control measures.

Longitudinal retrospective study on the effect of season on milk production and composition in Rio Grande do Sul, Brazil / Estudo retrospectivo longitudinal sobre o efeito da estação na produção e composição do leite no Rio Grande do Sul, Brasil

This longitudinal retrospective study evaluated production and composition of milk from dairy cows according to season on a rural property in Palmeira das Missões, Rio Grande do Sul, Brazil, between January 2009 and December 2016. Milk yield per hectare per month was the primary variable measured, along with chemical composition (total dry extract, defatted dry extract, fat, protein, and lactose), somatic cell count, and total bacterial count of milk. Production in summer (513.53 kg/ha/month) did not differ (p > 0.05) from that in fall (504.69 kg/ha/month) or spring (564.63 kg/ha/month), but production in winter (639.20 kg/ha/month) was significantly higher than in summer (p < 0.0057). The real price of milk did not differ (p > 0.05) among seasons. The total dry extract content differed (p < 0.0059) between seasons. However, the cooler fall and winter seasons favored the production of total solids. The defatted dry extract content was higher (p < 0.0001) in winter (8.65%), compared to the other seasons of the year. Fat content did not differ (p > 0.05) among seasons. Protein content differed (p < 0.0200) between summer (3.01%) and winter (3.16%), but both seasons did not differ (p > 0.05) from fall (3.13%) or spring (3.05%). Winter lactose levels (4.52%) were higher (p < 0.0011) than during other seasons. The somatic cell count did not differ (p > 0.05) among seasons, demonstrating that the evaluated herd possesses standard mammary gland health. The total bacterial count did not differ (p > 0.05) among the seasons, demonstrating that the hygiene of the facilities remains constant. As expected, the temperature and humidity index varied (p < 0.0001) with the seasons, and the deleterious effects were more pronounced in summer and spring, as they are the hottest seasons in humid subtropical environments. The results emphasize that the organized management of milk production reduces the variation in milk composition, making it easier to maintain a consistent high milk quality and also generating extra revenue.(AU)

Este estudo retrospectivo longitudinal avaliou a produção e a composição do leite de vacas leiteiras de acordo com a estação em uma propriedade rural em Palmeira das Missões, Rio Grande do Sul, Brasil, entre janeiro de 2009 e dezembro de 2016. A produção de leite por hectare por mês foi a principal variável mensurada, juntamente com a composição química (extrato seco total, extrato seco desengordurado, gordura, proteína e lactose), contagem de células somáticas e contagem bacteriana total de leite. A produção no verão (513,53 kg/ha/mês) não diferiu (p > 0,05) daquela no outono (504,69 kg/ha/mês) ou primavera (564,63 kg/ha/mês), mas a produção no inverno (639,20 kg/ha/mês) foi significativamente maior que no verão (p < 0,0057). O preço real do leite não diferiu (p > 0,05) entre as estações. O teor total de extrato seco diferiu (p < 0,0059) entre as estações. No entanto, as estações mais frias do outono e inverno favoreceram a produção de sólidos totais. O teor de extrato seco desengordurado foi maior (p < 0,0001) no inverno (8,65%), em relação as demais estações do ano. O conteúdo de gordura não diferiu (p > 0,05) entre as estações. O conteúdo de proteína diferiu (p < 0,0200) entre o verão (3,01%) e o inverno (3,16%), mas as duas estações não diferiram (p > 0,05) do outono (3,13%) ou da primavera (3,05%). Os níveis de lactose no inverno (4,52%) foram maiores (p < 0,0011) do que nas demais estações do ano. A contagem de células somáticas não diferiu (p > 0,05) entre as estações, demonstrando que o rebanho avaliado possui saúde padrão das glândulas mamárias. A contagem bacteriana total não diferiu (p > 0,05) entre as estações, demonstrando que a higiene das instalações permanece constante. Como esperado, o índice de temperatura e umidade variou (p < 0,0001) com as estações do ano e os efeitos deletérios foram mais pronunciados no verão e na primavera, por serem as estações mais quentes em ambientes subtropicais úmidos. Os resultados enfatizam que o manejo organizado da produção de leite reduz a variação na composição do leite, facilitando a manutenção de uma alta qualidade consistente do leite e também gerando receita extra.(AU)

Longitudinal Analysis of the Microbiological Quality of Raw Cow's Milk Samples Collected from Three Small Family Dairy Farms in Mexico over a 2-Year Period.

In Mexico, the total milk production that family dairy farms (FDF) contribute is ca. 35%, but this milk is not evaluated for microbiological quality. Forty percent of the milk and dairy products consumed by Mexicans is unpasteurized. In total, 24 raw cow's milk samples from three FDF (one sample per each season from each FDF for two sequent years) were characterized for the presence of food quality indicator organisms, Staphylococcus aureus, Salmonella enterica, Listeria monocytogenes, and Mycobacterium spp., by standard procedures. Escherichia coli presence was also evaluated by a direct count method and diarrheagenic E. coli (DEC) by molecular methods. On the basis of Mexican guidelines for raw milk entering production, 42% of samples exceeded the aerobic mesophilic bacteria limits. A total of 83% raw milk samples were positive for total coliforms, 54% for fecal coliforms, and 46% for E. coli. Forty-three E. coli isolates were selected and characterized for the presence of 11 DEC loci; of theses, 40 isolates were negative for all DEC loci, and 3 isolates, all collected from the same sample, were Shiga toxin 2 (stx2) positive and O157 antigen negative, and one stx2 isolate was resistant to 6 of the 16 antibiotics tested. None of the 24 raw milk samples were positive for Salmonella enterica, L. monocytogenes, or staphylococcal enterotoxin. S. aureus was isolated from nine samples, of which only three samples harbored resistant isolates. From three samples, four nontuberculous mycobacterial isolates were recovered (Mycobacteroides chelonae, Mycobacteroides porcinum, and two Mycobacteroides abscessus). All four isolates produced biofilm and had sliding motility, and three isolates (M. porcinum and two M. abscessus) were resistant to the two antibiotics tested (clarithromycin and linezolid). FDF provide raw milk to a large proportion of the Mexican population, but its consumption could be harmful to health, emphasizing the need to implement national microbiological quality guidelines for raw milk intended for direct human consumption.

Seroprevalence and Risk Factors Possibly Associated with Emerging Zoonotic Vaccinia Virus in a Farming Community, Colombia.

In 2014, vaccinia virus (VACV) infections were identified among farmworkers in Caquetá Department, Colombia; additional cases were identified in Cundinamarca Department in 2015. VACV, an orthopoxvirus (OPXV) used in the smallpox vaccine, has caused sporadic bovine and human outbreaks in countries such as Brazil and India. In response to the emergence of this disease in Colombia, we surveyed and collected blood from 134 farmworkers and household members from 56 farms in Cundinamarca Department. We tested serum samples for OPXV antibodies and correlated risk factors with seropositivity by using multivariate analyses. Fifty-two percent of farmworkers had OPXV antibodies; this percentage decreased to 31% when we excluded persons who would have been eligible for smallpox vaccination. The major risk factors for seropositivity were municipality, age, smallpox vaccination scar, duration of time working on a farm, and animals having vaccinia-like lesions. This investigation provides evidence for possible emergence of VACV as a zoonosis in South America.

Bovine Leukemia Virus Infection in Neonatal Calves. Risk Factors and Control Measures.

Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). Although efficient eradication programs have been successfully implemented in most European countries and Oceania, BLV infection rates are still high worldwide. BLV naturally infects cattle, inducing a persistent infection with diverse clinical outcomes. The virus infects lymphocytes and integrates a DNA intermediate as a provirus into the genome of the cells. Therefore, exposure to biological fluids contaminated with infected lymphocytes potentially spreads the virus. Vertical transmission may occur in utero or during delivery, and about 10% of calves born to BLV-infected dams are already infected at birth. Most frequently, transmission from dams to their offspring occurs through the ingestion of infected colostrum or milk. Therefore, although EBL is not a disease specific to the neonatal period, during this period the calves are at special risk of becoming infected, especially in dairy farms, where they ingest colostrum and/or raw milk either naturally or artificially. Calves infected during the first week of life could play an active role in early propagation of BLV to susceptible animals. This review discusses the main factors that contribute to neonatal BLV infection in dairy herds, as well as different approaches and management practices that could be implemented to reduce the risk of BLV transmission during this period, aiming to decrease BLV infection in dairy herds.

Detection of Listeria spp. in cattle and environment of pasture-based dairy farms / Detecção de Listeria spp. em gados leiteiros no meio ambiente com base na pastagem

The aim of the study was to detect Listeria spp., particularly Listeria monocytogenes, in cattle and environment of pasture based dairy farms in Paysandú, Uruguay. A two-stage sampling was conducted, 10 farms were selected by probability proportional to size. A single visit was made to each farm. Samples from bovine faeces, feedstuffs, bulk tank milk, drinking water and soil from the entry and exit pens of the milking parlour were collected for bacteriological studies. PCR assays were used to confirm species and determine the serotype profile of L. monocytogenes isolates. AscI-pulsed-field gel electrophoresis was done to genetically compare them. Listeria spp. were isolated from eight of ten dairy farms, whereas L. monocytogenes in three of them. Serotype distribution in L. monocytogenes was as follows: 1/2a, three isolates; 4b, one isolate. L. monocytogenes or L. innocua excreted from clinically healthy milking cows was detected via faeces. In feedstuffs, only one L. monocytogenes 1/2a isolate from a pasture was obtained. The strain was identical by PFGE to an isolate 1/2a obtained from a pool of milking cow feces that grazed on this farm. No isolation of Listeria spp. was retrieved from the bulk tank milk or drinking water from any of the farms. Listeria innocua was detected in 13 feedstuffs and seven samples of soil from the entry and exit pens of the milking parlour. This is a first local study that confirms the presence of Listeria spp. including L. monocytogenes in healthy cattle and environment of pasture-based dairy farms. These results suggest the potential role that healthy cattle and their sub-products would play as a source of these agents for humans and/or others animals. More detailed studies that include genetic comparison of human and animal isolates are required in order to clearly establish the epidemiological relationship.(AU)

O objetivo deste trabalho foi detectar a presença de bactérias do gênero Listeria e particularmente Listeria monocytogenes, em bovinos leiteiros no ambiente de Paysandú, Uruguai. Foi realizada uma amostragem em duas etapas, dez estabelecimentos foram selecionados por probabilidade proporcional ao tamanho. Foi realizada uma única visita a cada propriedade. Foram coletadas amostras para cultura bacteriológica de matéria fecal bovina, além de alimentos, leite do tanque de resfriamento, água e solo na entrada e saída da sala de ordenha. Com os isolados de L. monocytogenes foi realizado PCR para a confirmação da espécie e determinação do perfil do serotipo. AscI-elctroforese em gel de campo pulsado foi realizado para compará-los geneticamente. Listeria spp. foram isoladas de oito de dez estabelecimentos, enquanto L. monocytogenes foram detectadas em três deles. A distribuição dos serotipos nos isolados de L. monocytogenes recuperados foi: 1/2a três isolados, 4b um isolado. Foram detectadas vacas leiteras clinicamente sadias ​​que excretaram L. monocytogenes ou L. innocua nas fezes. Dos alimentos do gado houve só um isolamento de L. monocytogenes 1/2a em uma pastagem. Esta estirpe foi idêntica no PFGE a um isolado 1/2a obtido de uma "piscina" de fezes de vacas leiteiras do mesmo estabelecimento. Não houve isolamento de Listeria no leite do tanque de resfriamento ou na água de nenhum dos estabelecimentos. Listeria innocua foi detectada em 13 alimentos para o gado e sete amostras de solo na entrada e saída da sala de ordenha. Este parece ser o primeiro estudo local que confirma a presença de Listeria spp. incluindo L. monocytogenes em vacas leiteiras sadias e no meio ambiente de propriedades leiteiras com base alimentícia na pastagem. Esses resultados sugerem o potencial de vacas sadias e seus subprodutos como possível fonte desses agentes para humanos e/ou outros animais. São necessários estudos mais detalhados que incluem a comparação genética de isolados humanos e de animais para estabelecer claramente seu relacionamento epidemiológico.(AU)

Detection of Listeria spp. in cattle and environment of pasture-based dairy farms / Detecção de Listeria spp. em gados leiteiros no meio ambiente com base na pastagem

The aim of the study was to detect Listeria spp., particularly Listeria monocytogenes, in cattle and environment of pasture based dairy farms in Paysandú, Uruguay. A two-stage sampling was conducted, 10 farms were selected by probability proportional to size. A single visit was made to each farm. Samples from bovine faeces, feedstuffs, bulk tank milk, drinking water and soil from the entry and exit pens of the milking parlour were collected for bacteriological studies. PCR assays were used to confirm species and determine the serotype profile of L. monocytogenes isolates. AscI-pulsed-field gel electrophoresis was done to genetically compare them. Listeria spp. were isolated from eight of ten dairy farms, whereas L. monocytogenes in three of them. Serotype distribution in L. monocytogenes was as follows: 1/2a, three isolates; 4b, one isolate. L. monocytogenes or L. innocua excreted from clinically healthy milking cows was detected via faeces. In feedstuffs, only one L. monocytogenes 1/2a isolate from a pasture was obtained. The strain was identical by PFGE to an isolate 1/2a obtained from a pool of milking cow feces that grazed on this farm. No isolation of Listeria spp. was retrieved from the bulk tank milk or drinking water from any of the farms. Listeria innocua was detected in 13 feedstuffs and seven samples of soil from the entry and exit pens of the milking parlour. This is a first local study that confirms the presence of Listeria spp. including L. monocytogenes in healthy cattle and environment of pasture-based dairy farms. These results suggest the potential role that healthy cattle and their sub-products would play as a source of these agents for humans and/or others animals. More detailed studies that include genetic comparison of human and animal isolates are required in order to clearly establish the epidemiological relationship.(AU)

O objetivo deste trabalho foi detectar a presença de bactérias do gênero Listeria e particularmente Listeria monocytogenes, em bovinos leiteiros no ambiente de Paysandú, Uruguai. Foi realizada uma amostragem em duas etapas, dez estabelecimentos foram selecionados por probabilidade proporcional ao tamanho. Foi realizada uma única visita a cada propriedade. Foram coletadas amostras para cultura bacteriológica de matéria fecal bovina, além de alimentos, leite do tanque de resfriamento, água e solo na entrada e saída da sala de ordenha. Com os isolados de L. monocytogenes foi realizado PCR para a confirmação da espécie e determinação do perfil do serotipo. AscI-elctroforese em gel de campo pulsado foi realizado para compará-los geneticamente. Listeria spp. foram isoladas de oito de dez estabelecimentos, enquanto L. monocytogenes foram detectadas em três deles. A distribuição dos serotipos nos isolados de L. monocytogenes recuperados foi: 1/2a três isolados, 4b um isolado. Foram detectadas vacas leiteras clinicamente sadias ​​que excretaram L. monocytogenes ou L. innocua nas fezes. Dos alimentos do gado houve só um isolamento de L. monocytogenes 1/2a em uma pastagem. Esta estirpe foi idêntica no PFGE a um isolado 1/2a obtido de uma "piscina" de fezes de vacas leiteiras do mesmo estabelecimento. Não houve isolamento de Listeria no leite do tanque de resfriamento ou na água de nenhum dos estabelecimentos. Listeria innocua foi detectada em 13 alimentos para o gado e sete amostras de solo na entrada e saída da sala de ordenha. Este parece ser o primeiro estudo local que confirma a presença de Listeria spp. incluindo L. monocytogenes em vacas leiteiras sadias e no meio ambiente de propriedades leiteiras com base alimentícia na pastagem. Esses resultados sugerem o potencial de vacas sadias e seus subprodutos como possível fonte desses agentes para humanos e/ou outros animais. São necessários estudos mais detalhados que incluem a comparação genética de isolados humanos e de animais para estabelecer claramente seu relacionamento epidemiológico.(AU)

Pseudomonas spp: contamination sources in bulk tanks of dairy farms / Pseudomonas spp: fontes de contaminação em tanques de expansão em fazendas leiteiras

This study focused on isolating Pseudomonas spp. during milking process in ten dairy farms with manual and mechanical milking systems during dry and rainy seasons, and evaluating DNA homology and patterns of distribution between isolates, in order to identify main sources of milk contamination by Pseudomonas spp. A total of 167 isolates of Pseudomonas spp. were obtained from water, milkers' hands, cows' teats, teat cups, cooling tanks and raw milk. Bacteria of Pseudomonas spp. genus were isolated from 85 and 82 sampling points in dairy farms with manual and mechanical milking system, respectively. A significant difference (p=0.02) on Pseudomonas spp. isolation was observed among samples of surface of cows' teats before and after pre-dipping, but no significant difference (p>0.05) was observed among milking systems or seasons. The possibility of the same Pseudomonas spp. patterns are distributed in different farms and seasons using Amplified Fragment Length Polymorphism (AFLP) technique was demonstrated. Milkers' hands, surface of cows' teats, teat cups and cooling tanks were associated with raw milk contamination with Pseudomonas spp. on farms with manual and mechanical milking system, showing that regardless of the type of milking system and season, proper hygiene procedures of equipment, utensils and workers' hands are essential to avoid contamination of the milk and, therefore, improve milk quality.(AU)

Este estudo se propôs a isolar Pseudomonas spp. durante o processo de ordenha em dez fazendas com sistemas manuais e mecanizados, durante as estações seca e chuvosa, além de avaliar a homologia do DNA e seus padrões de distribuição entre os isolados, a fim de se determinar as principais fontes de contaminação do leite. Cento e sessenta e sete isolados de Pseudomonas spp. foram obtidos a partir de amostras de água, mãos de ordenhadores, tetos, teteiras, tanques de resfriamento e leite cru armazenado, sendo 85 e 82 pontos de amostragem em fazendas com sistemas de ordenha manual e mecânico, respectivamente. Diferença estatisticamente significativa foi encontrada entre os isolados observados entre a superfície dos tetos antes e após o pré-dipping (p=0,02), mas nenhuma diferença foi encontrada entre sistemas de ordenha ou estações (p>0,05). A possibilidade do mesmo padrão de Pseudomonas spp. estar distribuído em diferentes fazendas ou estações foi avaliada pela técnica de Polimorfismo do Tamanho de Fragmento Amplificado (AFLP). As mãos de ordenhadores, superfície dos tetos das vacas, teteiras e tanques de resfriamento foram associados com a contaminação do leite cru, demonstrando que independente do tipo de ordenha e estação, a higiene adequada de equipamentos, utensílios e mãos dos ordenhadores é essencial para evitar contaminação do leite, e consequentemente aumentar sua qualidade.(AU)

Pseudomonas spp: contamination sources in bulk tanks of dairy farms / Pseudomonas spp: fontes de contaminação em tanques de expansão em fazendas leiteiras

This study focused on isolating Pseudomonas spp. during milking process in ten dairy farms with manual and mechanical milking systems during dry and rainy seasons, and evaluating DNA homology and patterns of distribution between isolates, in order to identify main sources of milk contamination by Pseudomonas spp. A total of 167 isolates of Pseudomonas spp. were obtained from water, milkers' hands, cows' teats, teat cups, cooling tanks and raw milk. Bacteria of Pseudomonas spp. genus were isolated from 85 and 82 sampling points in dairy farms with manual and mechanical milking system, respectively. A significant difference (p=0.02) on Pseudomonas spp. isolation was observed among samples of surface of cows' teats before and after pre-dipping, but no significant difference (p>0.05) was observed among milking systems or seasons. The possibility of the same Pseudomonas spp. patterns are distributed in different farms and seasons using Amplified Fragment Length Polymorphism (AFLP) technique was demonstrated. Milkers' hands, surface of cows' teats, teat cups and cooling tanks were associated with raw milk contamination with Pseudomonas spp. on farms with manual and mechanical milking system, showing that regardless of the type of milking system and season, proper hygiene procedures of equipment, utensils and workers' hands are essential to avoid contamination of the milk and, therefore, improve milk quality.(AU)

Este estudo se propôs a isolar Pseudomonas spp. durante o processo de ordenha em dez fazendas com sistemas manuais e mecanizados, durante as estações seca e chuvosa, além de avaliar a homologia do DNA e seus padrões de distribuição entre os isolados, a fim de se determinar as principais fontes de contaminação do leite. Cento e sessenta e sete isolados de Pseudomonas spp. foram obtidos a partir de amostras de água, mãos de ordenhadores, tetos, teteiras, tanques de resfriamento e leite cru armazenado, sendo 85 e 82 pontos de amostragem em fazendas com sistemas de ordenha manual e mecânico, respectivamente. Diferença estatisticamente significativa foi encontrada entre os isolados observados entre a superfície dos tetos antes e após o pré-dipping (p=0,02), mas nenhuma diferença foi encontrada entre sistemas de ordenha ou estações (p>0,05). A possibilidade do mesmo padrão de Pseudomonas spp. estar distribuído em diferentes fazendas ou estações foi avaliada pela técnica de Polimorfismo do Tamanho de Fragmento Amplificado (AFLP). As mãos de ordenhadores, superfície dos tetos das vacas, teteiras e tanques de resfriamento foram associados com a contaminação do leite cru, demonstrando que independente do tipo de ordenha e estação, a higiene adequada de equipamentos, utensílios e mãos dos ordenhadores é essencial para evitar contaminação do leite, e consequentemente aumentar sua qualidade.(AU)

Molecular analysis of a fragment of bovine leukemia virus env gene by Nested-PCR in dairy cows from Pasto, Nariño / Análisis molecular de un fragmento del gen env del virus de leucosis bovina, por PCR anidada en vacas lecheras de Pasto, Nariño / Análise molecular pela reação em cadeia da polimerase de um fragmento do gene env do vírus de leucose bovina aninhada em vacas leiteiras de Pasto, Nariño

Abstract: Introduction: Routine diagnosis for bovine leukemia is performed using indirect serologic tests, although it is recommended to use polymerase chain reaction (PCR), which allows a reliable diagnosis in early stages and in young animals. Moreover, by amplifying the DNA, it is possible to sequence fragments of the virus, which allows to identify genotypes and to construct phylogenetic trees. Objective: To analyze a fragment of the env gene of bovine leukemia virus (BLV) isolated from indirect ELISA-positive animals in dairy farms in the municipality of Pasto (Nariño). Materials and methods: Once the presence of BLV was established in 48 animals over two years old in seven dairy farms in the municipality of Pasto by indirect ELISA test, a nested PCR test was performed to confirm diagnosis and to sequence a fragment of the env gene in positive animals and their daughters. The sequences obtained were compared with the seven genotypes described worldwide by MEGA 6 program. Results: The sequences of the compared fragments do not differ from those described; but they allow their grouping into different clusters according to their similarity. The genotypes found in these farms correspond to genotype 1 and 2 described in the literature. Conclusions: Only one genotype was found on farms with proper recovery and adequate biosecurity measures, and on farms with less control in management and recovery practices, both genotypes described for the region were evident.

Resumen: Introducción: El diagnóstico de rutina para la leucosis bovina se realiza con pruebas serológicas indirectas, aunque se recomienda usar la reacción en cadena de la polimerasa (PCR), que permite un diagnóstico confiable en fases iniciales y en animales jóvenes. Además, mediante la amplificación del ADN se pueden secuenciar fragmentos del virus, que permite identificar los genotipos presentes y construir árboles filogenéticos. Objetivo: Analizar un fragmento del gen env del virus de leucosis bovina (VLB) aislado de animales positivos a ELISA indirecta en fincas lecheras del municipio de Pasto, Nariño. Materiales y métodos: Una vez establecida la presencia del VLB en 48 animales mayores de dos años en siete fincas lecheras del municipio de Pasto mediante la prueba de ELISA indirecta, se realizó una prueba de PCR anidada para confirmar el diagnóstico y secuenciar un fragmento del gen env en los animales positivos y sus hijas. Las secuencias obtenidas se compararon con los siete genotipos descritos en el mundo mediante el programa MEGA 6. Resultados: Las secuencias de los fragmentos comparados no difieren de las descritas; pero permiten su agrupación en diferentes clústeres de acuerdo con su similitud. Los genotipos encontrados en estas fincas corresponden al genotipo 1 y 2 descritos en la literatura. Conclusiones: En las fincas con reposición propia y adecuadas medidas de bioseguridad solo se encontró un genotipo y en las fincas con menor control en las prácticas de manejo y de reposición se encontraron los dos genotipos descritos para la región.

Resumo: Introdução: O diagnóstico de rotina para a leucose bovina se realiza com provas serológicas indiretas, mesmo que se recomenda usar a reação em cadeia da polimerase (PCR), que permite um diagnóstico confiável em fases iniciais e em animais jovens. Além do mais, mediante a amplificação do ADN se podem sequenciar fragmentos do vírus, que permite identificar os genótipos presentes e construir árvores filogenéticas. Objetivo: Analisar um fragmento do gene env do vírus de leucose bovina (VLB) isolado de animais positivos a ELISA indireta em fazendas leiteiras do município de Pasto, Nariño. Materiais e métodos: Uma vez estabelecida a presença do VLB em 48 animais maiores de dois anos em sete fazendas leiteiras do município de Pasto mediante a prova de ELISA indireta, se realizou uma prova de PCR aninhada para confirmar o diagnóstico e sequenciar um fragmento do gene env nos animais positivos e suas filhas. As sequências obtidas foram compararam com os sete genótipos descritos no mundo mediante o programa MEGA 6. Resultados: As sequências dos fragmentos comparados não diferem das descritas; mas permitem a sua agrupação em diferentes clusters de acordo com sua similitude. Os genótipos encontrados nestas fazendas correspondem ao genótipo 1 e 2 descritos na literatura. Conclusões: Nas fazendas com reposição própria e adequadas medidas de biossegurança somente se encontrou um genótipo e nas fazendas com menos controle nas práticas de manejo e de reposição se encontraram os dois genótipos descritos para a região.

Pseudomonas spp.: contamination sources in bulk tanks of dairy farms

ABSTRACT: This study focused on isolating Pseudomonas spp. during milking process in ten dairy farms with manual and mechanical milking systems during dry and rainy seasons, and evaluating DNA homology and patterns of distribution between isolates, in order to identify main sources of milk contamination by Pseudomonas spp. A total of 167 isolates of Pseudomonas spp. were obtained from water, milkers hands, cows teats, teat cups, cooling tanks and raw milk. Bacteria of Pseudomonas spp. genus were isolated from 85 and 82 sampling points in dairy farms with manual and mechanical milking system, respectively. A significant difference (p=0.02) on Pseudomonas spp. isolation was observed among samples of surface of cows teats before and after pre-dipping, but no significant difference (p>0.05) was observed among milking systems or seasons. The possibility of the same Pseudomonas spp. patterns are distributed in different farms and seasons using Amplified Fragment Length Polymorphism (AFLP) technique was demonstrated. Milkers hands, surface of cows teats, teat cups and cooling tanks were associated with raw milk contamination with Pseudomonas spp. on farms with manual and mechanical milking system, showing that regardless of the type of milking system and season, proper hygiene procedures of equipment, utensils and workers hands are essential to avoid contamination of the milk and, therefore, improve milk quality.

RESUMO: Este estudo se propôs a isolar Pseudomonas spp. durante o processo de ordenha em dez fazendas com sistemas manuais e mecanizados, durante as estações seca e chuvosa, além de avaliar a homologia do DNA e seus padrões de distribuição entre os isolados, a fim de se determinar as principais fontes de contaminação do leite. Cento e sessenta e sete isolados de Pseudomonas spp. foram obtidos a partir de amostras de água, mãos de ordenhadores, tetos, teteiras, tanques de resfriamento e leite cru armazenado, sendo 85 e 82 pontos de amostragem em fazendas com sistemas de ordenha manual e mecânico, respectivamente. Diferença estatisticamente significativa foi encontrada entre os isolados observados entre a superfície dos tetos antes e após o pré-dipping (p=0,02), mas nenhuma diferença foi encontrada entre sistemas de ordenha ou estações (p>0,05). A possibilidade do mesmo padrão de Pseudomonas spp. estar distribuído em diferentes fazendas ou estações foi avaliada pela técnica de Polimorfismo do Tamanho de Fragmento Amplificado (AFLP). As mãos de ordenhadores, superfície dos tetos das vacas, teteiras e tanques de resfriamento foram associados com a contaminação do leite cru, demonstrando que independente do tipo de ordenha e estação, a higiene adequada de equipamentos, utensílios e mãos dos ordenhadores é essencial para evitar contaminação do leite, e consequentemente aumentar sua qualidade.

Calidad de la leche y el perfil de los productores de una linea de transporte de la leche / Milk quality and profile of producers from a milk pick-up route / Qualidade do leite e perfil dos produtores de uma linha de entrega de leite

El objetivo de este estudio fue una evaluación secuencial de la calidad de la leche y el perfil de 34 productores de leche en la región central y un productor de la región de Campinas en Sao Paulo. Se recogieron los datos de las propiedades durante tres visitas con un intervalo promedio de dos meses, incluyendo un cuestionario para comprobar el tamaño de los rebaños y la gestión realizada. También se registraron los datos mensuales del recuento total de bacterias (TBC) y el recuento de células somáticas (SCC) proporcionada por la industria láctea. Los productores de la región tenían características diferentes como tres a 111 animales lactantes, variables de producción diaria de 10 a 3.632,8 litros. De las 35 propiedades, 15.4% y 27.89% tenían un recuento medio de células somáticas (SCC) y el recuento total de bacterias (TBC), respectivamente, por encima de los límites establecidos por el Ministerio de Agricultura, Ganadería y Abastecimiento (MAPA), la de acuerdo con la instrucción normativa 51, vigente en el momento. Sólo el 17,65% de las propiedades a cabo todos los procedimientos de higiene para el ordeño y el 17,65% no recibió ningún tipo de soporte técnico. Se observó que las propiedades son pequeñas a de tamaño medio, con el potencial para aumentar la producción y mejorar la calidad de la leche, como ya resfriavan el producto y tenía algún grado de tecnificación. [...]

The aim of this study was to perform a follow-up evaluation of both the milk quality and the profile of 34 dairy milk producers at the central-west region of São Paulo State and one producer at Campinas region, also in São Paulo State, Brazil. Data were collected from farms during three visits at two-month mean intervals, including a questionnaire to verify the herd size and the adopted management. Monthly data of total bacterial count (TBC) and somatic cell count (SCC) provided by the dairy plant were also recorded. The producers had variable characteristics such as: from three to 111 lactating animals; daily production ranging from 10 to 3,632.8 litters. Of the 35 farms, 15.4% and 27.8% had SCC and TBC, respectively above the limits established by the Brazilian Ministry of Agriculture Livestock and Food Supply (MAPA), according the Normative Instruction (IN 51). Only 17.65% farms carried out all hygienic procedures recommended for milking, while 17.65% do not received any type of technical support. The studied farms belong to small to medium-scale farmers with the potential to increase their production and improve milk quality, since they already refrigerated the product and had a certain automation degree. These results indicate the importance of adopting sanitary education activities for the production of more hygienic milk, as well as actions for mastitis control.

O objetivo deste estudo foi realizar uma avaliação sequencial da qualidade do leite e o perfil de 34 produtores de leite da região Centro-Oeste e de um produtor da região de Campinas no estado de São Paulo. Os dados das propriedades foram coletados durante três visitas com intervalo médio de dois meses, incluindo um questionário para verificar o tamanho do rebanho e manejo realizado. Foram anotados ainda os dados mensais de contagem bacteriana total (CBT) e contagem de células somáticas (CCS) fornecidos pelo laticínio. Os produtores da região tinham características diferentes como: de três a 111 animais em lactação, produção diária variável de 10 a 3.632,8 litros. Das 35 propriedades, 15,4% e 27,89% apresentaram em média contagem de células somáticas (CCS) e contagem bacteriana total (CBT), respectivamente, acima dos limites determinados pelo Ministério da Agricultura, Pecuária e Abastecimento (MAPA), de acordo com a instrução normativa 51, vigente na época. Apenas 17,65% das propriedades realizavam todos os procedimentos de higiene na ordenha e 17,65% não recebiam nenhum tipo de suporte técnico. Foi possível observar que as propriedades são de produtores de pequeno a médio porte, com potencial para aumentar a produção e melhorar a qualidade do leite, pois já refrigeravam o produto e apresentavam algum grau de tecnificação. Conclui-se ainda pela importância da adoção de [...]

Calidad de la leche y el perfil de los productores de una linea de transporte de la leche / Milk quality and profile of producers from a milk pick-up route / Qualidade do leite e perfil dos produtores de uma linha de entrega de leite

El objetivo de este estudio fue una evaluación secuencial de la calidad de la leche y el perfil de 34 productores de leche en la región central y un productor de la región de Campinas en Sao Paulo. Se recogieron los datos de las propiedades durante tres visitas con un intervalo promedio de dos meses, incluyendo un cuestionario para comprobar el tamaño de los rebaños y la gestión realizada. También se registraron los datos mensuales del recuento total de bacterias (TBC) y el recuento de células somáticas (SCC) proporcionada por la industria láctea. Los productores de la región tenían características diferentes como tres a 111 animales lactantes, variables de producción diaria de 10 a 3.632,8 litros. De las 35 propiedades, 15.4% y 27.89% tenían un recuento medio de células somáticas (SCC) y el recuento total de bacterias (TBC), respectivamente, por encima de los límites establecidos por el Ministerio de Agricultura, Ganadería y Abastecimiento (MAPA), la de acuerdo con la instrucción normativa 51, vigente en el momento. Sólo el 17,65% de las propiedades a cabo todos los procedimientos de higiene para el ordeño y el 17,65% no recibió ningún tipo de soporte técnico. Se observó que las propiedades son pequeñas a de tamaño medio, con el potencial para aumentar la producción y mejorar la calidad de la leche, como ya resfriavan el producto y tenía algún grado de tecnificación. [...](AU)

The aim of this study was to perform a follow-up evaluation of both the milk quality and the profile of 34 dairy milk producers at the central-west region of São Paulo State and one producer at Campinas region, also in São Paulo State, Brazil. Data were collected from farms during three visits at two-month mean intervals, including a questionnaire to verify the herd size and the adopted management. Monthly data of total bacterial count (TBC) and somatic cell count (SCC) provided by the dairy plant were also recorded. The producers had variable characteristics such as: from three to 111 lactating animals; daily production ranging from 10 to 3,632.8 litters. Of the 35 farms, 15.4% and 27.8% had SCC and TBC, respectively above the limits established by the Brazilian Ministry of Agriculture Livestock and Food Supply (MAPA), according the Normative Instruction (IN 51). Only 17.65% farms carried out all hygienic procedures recommended for milking, while 17.65% do not received any type of technical support. The studied farms belong to small to medium-scale farmers with the potential to increase their production and improve milk quality, since they already refrigerated the product and had a certain automation degree. These results indicate the importance of adopting sanitary education activities for the production of more hygienic milk, as well as actions for mastitis control.(AU)

O objetivo deste estudo foi realizar uma avaliação sequencial da qualidade do leite e o perfil de 34 produtores de leite da região Centro-Oeste e de um produtor da região de Campinas no estado de São Paulo. Os dados das propriedades foram coletados durante três visitas com intervalo médio de dois meses, incluindo um questionário para verificar o tamanho do rebanho e manejo realizado. Foram anotados ainda os dados mensais de contagem bacteriana total (CBT) e contagem de células somáticas (CCS) fornecidos pelo laticínio. Os produtores da região tinham características diferentes como: de três a 111 animais em lactação, produção diária variável de 10 a 3.632,8 litros. Das 35 propriedades, 15,4% e 27,89% apresentaram em média contagem de células somáticas (CCS) e contagem bacteriana total (CBT), respectivamente, acima dos limites determinados pelo Ministério da Agricultura, Pecuária e Abastecimento (MAPA), de acordo com a instrução normativa 51, vigente na época. Apenas 17,65% das propriedades realizavam todos os procedimentos de higiene na ordenha e 17,65% não recebiam nenhum tipo de suporte técnico. Foi possível observar que as propriedades são de produtores de pequeno a médio porte, com potencial para aumentar a produção e melhorar a qualidade do leite, pois já refrigeravam o produto e apresentavam algum grau de tecnificação. Conclui-se ainda pela importância da adoção de [...](AU)