Periparturient blood T-lymphocyte PD-1 and CTLA-4 expression as potential predictors of new intramammary infections in dairy cows during early lactation (short communication).

BACKGROUND: The periparturient period in dairy cows is marked by immunosuppression which increases the likelihood of infectious disorders, particularly also mastitis. An in-depth understanding of peripartum leukocyte biology is vital for the implementation of highly successful post-partum disease prevention measures. Immune checkpoint molecules, such as programmed death 1 (PD-1) and cytotoxic T-lymphocyte antigen 4 (CTLA-4), are critical inhibitory receptors expressed on immune cells, particularly T cells, that drive immunosuppressive signaling pathways. However, the potential role of immune checkpoint molecules expression in T-cells on udder health has never been explored. Thus, the association between the occurrence of new postpartum intramammary infections (IMIs) and the expression of programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) on blood T-cells during the peripartum period was investigated. RESULTS: In this study, the incidence of IMIs by any pathogen in early lactation was not associated with a higher expression of PD-1 and CTLA-4 in the periparturient period. However, the incidence of IMIs by major pathogens throughout the first month of lactation was significantly associated with higher expression of PD-1 at 14 days before calving (P = 0.03) and CTLA-4 at parturition (P = 0.03) by blood T-cells. Also, the expression of CTLA-4 at D0 (P = 0.012) by T-cells was associated with the occurrence of persistent IMIs during the first month of lactation. CONCLUSIONS: To our knowledge, this is the first report to investigate the expression of PD-1 and CTLA-4 by blood T-lymphocytes during the periparturient period in dairy cows and to explore their relationship with the incidence of new IMIs in the postpartum period. Thus, a comprehensive understanding of leukocyte biology during peripartum would appear to be a prerequisite for the identification of resilient dairy cows or targets innovative (immunological) non-antibiotic approaches in the transition period.

Action of swim-up and caffeine on equine frozen sperm.

Cryopreservation of equine semen is crucial to semen commercialization. However, it reduces sperm motility and longevity. Thus, sperm selection methods and addition of motility-activating substances to sperm, such as caffeine, may improve sperm quality of equine frozen semen. The objective of the current work was to evaluate the effects of caffeine on recovery and quality parameters of frozen-thawed sperm subjected to swim-up selection to be used in intracytoplasmic sperm injection (ICSI) in assisted reproductive techniques. Stallion semen were frozen and after thawing different caffeine concentrations were added to the samples performing four treatments control (no caffeine), 3, 5, and 7.5 mM caffeine. Sperm kinematic and motility were assessed by computer-assisted sperm analysis (CASA). Then, the four treated samples were submitted to the swim-up sperm selection, and the number of recovered sperm and morphology were evaluated at four times 20, 40, 60, and 80 min. The swim-up increased the recovery proportion of normal morphology sperm without (80.1±1%) or with caffeine addition (3mM: 81.2±1%, 5mM: 79.9±1% and 7.5 mM 78.9±1%) compared to the thawed semen (70±2%). However, the addition of 5 mM caffeine induced an increase in sperm motility (38.9±2.8 vs. 32.6±3.4%, P<0.05), and sperm recovery after swim-up (7.9x106 vs. 3.4x106 sperm/ml, P<0.05) compared to the control. The addition of 5 mM caffeine to frozen-thawed equine semen before swim-up selection improved sperm motility and increased the sperm recovery rate while not decreasing the percentage of morphologically normal sperm. Thus, caffeine addition to frozen-thawed equine semen before swim-up selection has potential clinical application in improving sperm quality for use in ICSI.

Milk Macrophage Function in Bovine Leukemia Virus-Infected Dairy Cows.

The implications of bovine leukemia virus (BLV) on innate and adaptive immune responses have been widely investigated; however, the effects of BLV on mammary gland immunity require further investigation. The present study investigated the viability, phagocytic capacity, and intracellular production of reactive oxygen and nitrogen species (RONS) by macrophages in milk samples from dairy cows naturally infected with BLV with or without persistent lymphocytosis (PL). No effect of BLV infection in the overall number of macrophages per milliliter and in the percentage of viable macrophages among overall milk viable cells was found. Furthermore, BLV-infected dairy cows had a higher frequency of viable milk macrophages, while healthy animals had a tendency toward a higher percentage of apoptotic milk macrophages. The percentage of milk macrophages that phagocytosed Staphylococcus aureus in seronegative animals was higher than that in BLV-infected dairy cows. No effect of BLV infection on the intracellular RONS production and the intensity of phagocytosis by milk macrophages was observed. Thus, this study provides new insights into the implications of BLV infections in the bovine mammary gland.

Spatial distribution of anti-rabies post-exposure treatments associated with the decentralization process in Belo Horizonte, 2007 to 2011

Cases of canine attacks on people are reported because of the proximity of the dog to the households in several Brazilian cities. In the present study, we aim to evaluate post-exposure anti-rabies treatments with canine accidents between the years 2007 to 2011 in Belo Horizonte - MG. Duly notified data were obtained from the National System of Notifiable Diseases (SINAN). The spatial characteristics of the cases during the period of the study referred to the neighborhoods and the nine sanitary districts of the municipality of Belo Horizonte - MG. For georeferencing and spatial analysis, we used the software Maporama to identify the coordinates and the Geographic Information System ArcGIS for mapping. Considering the 6.153 prophylactic services that were georeferenced, the Norte, Venda Nova, Leste, and Centro Sul regions were highlighted due to the higher frequency of registered cases. It is suggested that an integrative action focused on canine population control, health education and epidemiological surveillance studies could contribute to the reduction of canine aggression cases.

Addition of caffeine to equine thawed sperm increases motility and decreases nitrite concentration.

The aim of this study was to improve the quality of frozen-thawed equine sperm by the addition of caffeine to it. Semen from nine stallions was frozen and different concentrations of caffeine (3, 5 and 7.5 mM) were added to frozen-thawed semen. The sperm kinetic parameters, membrane functionality and integrity, and acrosome integrity and spontaneous acrosome reacted sperm were evaluated with a computer-assisted sperm analysis, a hypoosmotic swelling test and epifluorescent microscopy, respectively. Nitrite and hydroperoxide concentrations of frozen-thawed semen were measured using spectrophotometry. Sperm fertility was evaluated by artificial insemination (AI) of 16 mares with thawed ejaculates (control and 5 mM caffeine-treated groups). Compared to that in the control, the addition of 5 mM caffeine induced an increase in sperm motility (38.9 ± 2.8 versus 32.6 ± 3.4%), and a decrease in nitrite concentration (11.4 ± 2.1 versus 12.8 ± 2.9 µM/µg protein, p < .05). Moreover, the pregnancy rate from AI in the caffeine group was significantly higher (62.5%) than that in the control group (12.5%). These data suggest that caffeine reduced the nitrite concentration and enhanced sperm motility in thawed equine sperm, thus increasing the fertility rate in mares inseminated with caffeine-treated equine semen.

Profile of American tegumentary leishmaniasis in transmission areas in the state of Minas Gerais, Brazil, from 2007 to 2017.

BACKGROUND: American tegumentary leishmaniasis (ATL) is a widespread anthropozoonosis caused by protozoa of the genus Leishmania and is considered a serious public health problem. The aim of this study was to provide a descriptive analysis of confirmed ATL cases and evaluate the spatial distribution of ATL in high-risk transmission areas from the state of Minas Gerais, Brazil. METHODS: An ecological, analytical, and retrospective study of the confirmed cases of ATL in Minas Gerais from 2007 to 2017 was conducted. To characterize these cases, multiple correspondence analysis and georeferencing of the ATL prevalence rates in the municipalities were conducted based on variables obtained at Sistema Nacional de Agravos de Notificação and Instituto Brasileiro de Geografia e Estatística databases. RESULTS: There were 13,025 confirmed cases of ATL from 74.4% (635) municipalities of Minas Gerais, corresponding to a prevalence rate of 66.5 cases for every 100,000 inhabitants. Males aged 20 to 59 years and individuals who attended elementary school were most affected with ATL. Multiple correspondence analysis presented an accumulated qui-squared value of 44.74%, proving that there was a relationship between the variables, including ethnicity, age, pregnancy status, zone of infection, and number of cases. CONCLUSION: We confirmed that ATL is endemic to Minas Gerais, and there is high risk of infection within the municipalities due to a high rate of parasite transmission. The occurrence of infection in children, pregnant women, and the indigenous population demonstrates the need for the government to expand social policies aimed at vulnerable groups.

Spatio-temporal study of diagnosis of rabies in vampire bats in Sergipe (Brazil), between 1987 and 2014 / Estudo espaço-temporal de diagnósticos da raiva em morcegos hematófagos em sergipe (brasil), entre 1987 e 2014

Rabies is a zoonosis of great importance for human and animal health, being responsible for a progressive and acute encephalitis in humans and animals. Considering that the correct viral identification has serious implications for the prophylaxis and treatment, is important a diagnostic system specifically designed for monitoring the infection cases. The objective of the study was to describe the occurrence of rabies in hematophagous bats (Desmodus rotundus) analyzing many diagnoses performed in the state of Sergipe. The objective of this study was to describe the occurrence of rabies in hematophagous bats (Desmodus rotundus) from an analysis of the diagnoses performed in the state of Sergipe. An observational, descriptive and retrospective study was conducted with the evaluation of 935 rabies diagnoses in bats (D. rotundus) between 1987 and 2014. It was noticed that only one case was positive in the city of Aracaju in the year 2000 and no positive bat was identified with the rabies virus in the rural area. The results show the need to increase the monitoring of rabies in other species of non-hematophagous bats, and especially in other wild reservoirs. It is suggested that the monitoring of rabies take into account the specific ecosystem of each region, since Brazil has continental dimensions and a great biological diversity.

Araiva é uma zoonose de grande importância para a saúde humana e animal, sendo responsável por uma encefalite progressiva e aguda em humanos e animais. Considerando que a correta identificação viral tem sérias implicações para a profilaxia e tratamento, é importante um sistema de diagnóstico especificamente projetado para monitorar os casos de infecção. O estudo teve como objetivo descrever a ocorrência da raiva em morcegos hematófagos (Desmodus rotundus) a partir da análise dos diagnósticos realizados no estado de Sergipe. Foi realizado um estudo observacional, descritivo e retrospectivo com avaliação de 935 diagnósticos de raiva em morcegos (D. rotundus) entre os anos de 1987 a 2014. Percebeu-se que apenas um caso foi positivo na cidade de Aracaju no ano 2000 e nenhum morcego positivo foi identificado com o vírus da raiva na área rural. Os resultados demonstram a necessidade de ampliar o monitoramento da raiva em outras espécies de morcegos não hematófagos e sobretudo em outros reservatórios selvagens. Sugere-se que o monitoramento da raiva leve em consideração o ecossistema específico de cada região brasileira, já que o Brasil tem dimensões continentais e uma grande diversidade biológica.

Epidemiological characterization and risk evaluation associated with the presence of Phthirapterain poultry farms from Minas Gerais, Brazil

Phthirapteraare lice highly adapted to living as permanent and obligatory ectoparasites of birds and mammals. High infestations by biting lice contribute to the low productivity, loss quality of the eggs, besides weight decrease of chickens. Therefore, this study aimed to evaluate and to characterize the risks involved with the presence of biting lice in laying chicken from Minas Gerais, Brazil, throught a Multiple Correspondence Analysis (MCA). A secondary database with information regarding 402 chicken houses from 42 properties of laying hens from the same region was used and adapted for this study. The variables that composed the correspondence model were selected through Chi-square tests (p ≤ 0.05). A risk index was designed to relate the presence of biting lice out of variables such as risk factors related to the presence of these ectoparasites. The MCA graphic designed to conduct the epidemiological characterization of the presence of biting lice presented a Chi-square accumulated value of 53.59%. There was an association between the risk index and the synanthropic birds, subsistence hens, presence of lice and other ectoparasites in shelters and ectoparasites control. These results provide knowledge about the Phthirapteraepidemiology. Besides that, this information may contribute to the decision-making process in order to reduce the risk of possible infestations in poultry farms and the negative effects of the infestations.

Virulence factors and phylotyping of Escherichia coli isolated from non-diarrheic and diarrheic water buffalo calves / Caracterização dos fatores de virulência e filogrupos de Escherichia coli isoladas de bezerros bubalinos sem diarreia e diarréicos

ABSTRACT: This study aimed to determine the virulence factors, phylogenetic groups, and the relationships between pathovars and phylogenetic groups of E. coli strains isolated from feces of buffalo calves. A total of 217 E. coli strains were obtained from feces after culture and were screened by PCR for detection of virulence factors EAST-1, enterohemolysin, Saa, CNF2, F41, F5, STa, intimin, Stx1 and Stx2. One hundred and thirty-four isolates were positive for one or more virulence factors: eighty-four from diarrheic animals, and fifty from non-diarrheic calves. The pathovars of E. coli identified in diarrheic feces were ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), and EAEC (EAST-1+) (33/84). Pathovars identified in non-diarrheic animals were NTEC (21/50), STEC (17/50), EHEC (1/50) and EAEC (7/50). E. coli strains positive for EAST-1 (P=0.008) and phylogroup C (P = 0.05) were associated with the presence of diarrhea. Phylogenetic analysis showed that 58.95% of the isolates belonged to phylogroup B1, followed by E (9.70%), B2 (5.90%), C (5.90%), D (5.22%), A (2.24%), and F (1.50%). Phylogroup B1 predominated in pathogenic E. coli isolated from water buffalo, and phylogroup C constituted an enteropathogenic E. coli for water buffalo calves.

RESUMO: O objetivo foi determinar os fatores de virulência, os grupos filogenéticos e as possíveis relações entre os patovares e os grupos filogenéticos identificados de cepas de Escherichia coli isoladas de fezes de bezerros bubalinos. Um total de 217 amostras de E. coli foram identificadas a partir de cultura das fezes e submetidas a reação em cadeia da polimerase (PCR) para detecção dos fatores de virulência EAST-1, enterohemolisina, Saa, CNF2, F41, F5, STa, intimina, Stx1 e Stx2. Foram identificadas 134 cepas positivas para um ou mais fatores de virulência: 84isoladas de bezerros bubalinos diarreicos e 50 de bezerros bubalinos saudáveis. Os patovares de E. coli obtidos de fezes diarreicas foram ETEC (F5+) (2/84), NTEC (16/84), STEC (20/84), EPEC (3/84), EHEC (3/84), e EAEC (EAST-1+) (33/84). Os patovares isolados de fezes não diarreicas foram NTEC (21/50), STEC (17/50), EHEC (1/50) e EAEC (7/50). Cepas de E. coli positivas para EAST-1 (P = 0,008) e filogrupo C (P = 0,05) foram associadas com a presença de diarreia. A análise de filogrupos revelou que 58,95% dos isolados pertencem ao filogrupo B1, seguido por E (9,70%), B2 (5,90%), C (5,90%), D (5,22%), A (2,24%) e F (1,50%). O filogrupo B1 predomina em cepas de E. coli patogênicas isoladas de bezerros búfalos e o filogrupo C constitui um filogrupo de E. coli patogênica entérica para bezerros.

[American tegumentary leishmaniasis: a multivariate analysis of the spatial circuits for production of cases in Minas Gerais State, Brazil, 2007 to 2011]. / Ocorrência de casos de leishmaniose tegumentar americana: uma análise multivariada dos circuitos espaciais de produção, Minas Gerais, Brasil, 2007 a 2011.

The aim of this study was to explain the association between social-environmental factors and major land uses and the occurrence of cases of American tegumentary leishmaniasis (ATL) in the spatial circuits of production in Minas Gerais State, Brazil. This was an analytical-type ecological study based on secondary data on ATL divided by three-year period from 2007 to 2011, in which the analytical units were municipalities belonging to the spatial circuits. Two distinct stages were performed. The first was the elaboration of thematic maps with identification of the circuits. In the second, a new indicator, ATL cases by population density, was associated with social-environmental indicators and major land uses, submitted to multivariate principal components analysis (PCA). During the periods studied, three circuits were identified, distributed in the major regions of Northern Minas Gerais, Rio Doce Valley, and Greater Metropolitan Belo Horizonte. There was a strong association between ATL by population density and temporary crops, natural pasture, natural forest, unusable lands, and rural population, and a weak association with planted pasture. The association of cases with the major land uses variable in different agricultural profiles shows the occupational nature of ATL, associated mainly with rural workers. The association of the disease with environmental variables and deficient basic sanitation also proved relevant in the transmission profile in spatial circuits of production in Minas Gerais.

O objetivo deste estudo foi explicar a associação dos fatores socioambientais e dos grandes usos da terra com a ocorrência de casos de leishmaniose tegumentar americana (LTA) nos circuitos espaciais de produção, no Estado de Minas Gerais, Brasil. Trata-se de um estudo ecológico do tipo analítico, baseado em dados secundários de casos de LTA dividido por triênio, no período entre 2007 a 2011, cujas unidades de análise foram os municípios pertencentes aos circuitos espaciais. Duas etapas distintas foram realizadas, sendo a elaboração de mapas temáticos com a identificação dos circuitos a primeira, e na segunda etapa um novo indicador casos de LTA por densidade demográfica foi associado com indicadores socioambientais e dos grandes usos da terra submetidos à análise multivariada de componentes principais (ACP). Para o período avaliado, identificou-se três circuitos distribuídos nas mesorregiões Norte de Minas Gerais, Vale do Rio Doce e Região Metropolitana de Belo Horizonte. Houve forte associação dos casos de LTA por densidade demográfica com lavoura temporária, pastagem natural, floresta natural, terras inaproveitáveis e população rural, e uma fraca associação com pastagem plantada. A associação de casos com variáveis dos grandes usos da terra em diferentes perfis agropecuários demonstra o caráter ocupacional da LTA, associado principalmente com trabalhadores da zona rural. A associação da doença com as variáveis ambientais e deficiência das condições de saneamento básico também demonstram relevância no perfil de transmissão nos circuitos espaciais de produção em Minas Gerais.

El objetivo de este estudio fue explicar la asociación de los factores socioambientales y explotación a gran escala de la tierra, con la ocurrencia de casos de leishmaniasis tegumentaria americana (LTA) en los circuitos espaciales de producción, en el Estado de Minas Gerais, Brasil. Se trata de un estudio ecológico de tipo analítico, basado en datos secundarios de casos de LTA, dividido por trienios, en el período entre 2007 a 2011, cuyas unidades de análisis fueron los municipios pertenecientes a los circuitos espaciales. Se realizaron dos etapas distintas, consistiendo la primera en la elaboración de mapas temáticos con la identificación de los circuitos, y la segunda etapa en un nuevo indicador de casos de LTA por densidad demográfica, que se asoció con indicadores socioambientales y a dos explotaciones a gran escala de la tierra, sometidos a un análisis multivariado de componentes principales (ACP). Para el período evaluado, se identificaron tres circuitos distribuidos en las mesorregiones del norte de Minas Gerais, Vale do Rio Doce y región metropolitana de Belo Horizonte. Hubo una fuerte asociación de los casos de LTA por densidad demográfica con el trabajo temporal, pasto natural, bosque natural, tierras inaprovechables y población rural, y una escasa asociación con pasto plantado. La asociación de casos con variables de las explotaciones a gran escala de la tierra, con diferentes perfiles agropecuarios, demuestra el carácter ocupacional de la LTA, asociado principalmente a trabajadores de la zona rural. La asociación de la enfermedad con variables ambientales y deficiencia en las condiciones de saneamiento básico, también demuestran relevancia en el perfil de transmisión en los circuitos espaciales de producción en Minas Gerais.

Ocorrência de casos de leishmaniose tegumentar americana: uma análise multivariada dos circuitos espaciais de produção, Minas Gerais, Brasil, 2007 a 2011 / American tegumentary leishmaniasis: a multivariate analysis of the spatial circuits for production of cases in Minas Gerais State, Brazil, 2007 to 2011 / Ocurrencia de casos de leishmaniasis tegumentaria americana: un análisis multivariado de los circuitos espaciales de producción, Minas Gerais, Brasil, 2007 a 2011

O objetivo deste estudo foi explicar a associação dos fatores socioambientais e dos grandes usos da terra com a ocorrência de casos de leishmaniose tegumentar americana (LTA) nos circuitos espaciais de produção, no Estado de Minas Gerais, Brasil. Trata-se de um estudo ecológico do tipo analítico, baseado em dados secundários de casos de LTA dividido por triênio, no período entre 2007 a 2011, cujas unidades de análise foram os municípios pertencentes aos circuitos espaciais. Duas etapas distintas foram realizadas, sendo a elaboração de mapas temáticos com a identificação dos circuitos a primeira, e na segunda etapa um novo indicador casos de LTA por densidade demográfica foi associado com indicadores socioambientais e dos grandes usos da terra submetidos à análise multivariada de componentes principais (ACP). Para o período avaliado, identificou-se três circuitos distribuídos nas mesorregiões Norte de Minas Gerais, Vale do Rio Doce e Região Metropolitana de Belo Horizonte. Houve forte associação dos casos de LTA por densidade demográfica com lavoura temporária, pastagem natural, floresta natural, terras inaproveitáveis e população rural, e uma fraca associação com pastagem plantada. A associação de casos com variáveis dos grandes usos da terra em diferentes perfis agropecuários demonstra o caráter ocupacional da LTA, associado principalmente com trabalhadores da zona rural. A associação da doença com as variáveis ambientais e deficiência das condições de saneamento básico também demonstram relevância no perfil de transmissão nos circuitos espaciais de produção em Minas Gerais.

The aim of this study was to explain the association between social-environmental factors and major land uses and the occurrence of cases of American tegumentary leishmaniasis (ATL) in the spatial circuits of production in Minas Gerais State, Brazil. This was an analytical-type ecological study based on secondary data on ATL divided by three-year period from 2007 to 2011, in which the analytical units were municipalities belonging to the spatial circuits. Two distinct stages were performed. The first was the elaboration of thematic maps with identification of the circuits. In the second, a new indicator, ATL cases by population density, was associated with social-environmental indicators and major land uses, submitted to multivariate principal components analysis (PCA). During the periods studied, three circuits were identified, distributed in the major regions of Northern Minas Gerais, Rio Doce Valley, and Greater Metropolitan Belo Horizonte. There was a strong association between ATL by population density and temporary crops, natural pasture, natural forest, unusable lands, and rural population, and a weak association with planted pasture. The association of cases with the major land uses variable in different agricultural profiles shows the occupational nature of ATL, associated mainly with rural workers. The association of the disease with environmental variables and deficient basic sanitation also proved relevant in the transmission profile in spatial circuits of production in Minas Gerais.

El objetivo de este estudio fue explicar la asociación de los factores socioambientales y explotación a gran escala de la tierra, con la ocurrencia de casos de leishmaniasis tegumentaria americana (LTA) en los circuitos espaciales de producción, en el Estado de Minas Gerais, Brasil. Se trata de un estudio ecológico de tipo analítico, basado en datos secundarios de casos de LTA, dividido por trienios, en el período entre 2007 a 2011, cuyas unidades de análisis fueron los municipios pertenecientes a los circuitos espaciales. Se realizaron dos etapas distintas, consistiendo la primera en la elaboración de mapas temáticos con la identificación de los circuitos, y la segunda etapa en un nuevo indicador de casos de LTA por densidad demográfica, que se asoció con indicadores socioambientales y a dos explotaciones a gran escala de la tierra, sometidos a un análisis multivariado de componentes principales (ACP). Para el período evaluado, se identificaron tres circuitos distribuidos en las mesorregiones del norte de Minas Gerais, Vale do Rio Doce y región metropolitana de Belo Horizonte. Hubo una fuerte asociación de los casos de LTA por densidad demográfica con el trabajo temporal, pasto natural, bosque natural, tierras inaprovechables y población rural, y una escasa asociación con pasto plantado. La asociación de casos con variables de las explotaciones a gran escala de la tierra, con diferentes perfiles agropecuarios, demuestra el carácter ocupacional de la LTA, asociado principalmente a trabajadores de la zona rural. La asociación de la enfermedad con variables ambientales y deficiencia en las condiciones de saneamiento básico, también demuestran relevancia en el perfil de transmisión en los circuitos espaciales de producción en Minas Gerais.

Phylogenetic Group of Escherichia coli Isolates from Broilers in Brazilian Poultry Slaughterhouse.

The aim of the study was to determine the phylogenetic groups of E. coli strains isolated from seemingly healthy broiler and broiler condemned suspected of colibacillosis in a Brazilian slaughterhouse. Samples from respiratory tract and edible giblets (liver and heart) of broilers with and without macroscopic lesions of colibacillosis were collected at slaughter. There were 84 strains isolated from broilers condemned of which 11 were obtained from swabs of the heart, 7 from the liver, and 66 from the respiratory tract. Of the 53 E. coli strains isolated from broilers not condemned, 5 were isolated from the heart, 4 from the liver, and 44 from the respiratory tract. E coli strains were tested via PCR for phylogenetic groups A, B1, B2, C, D, E, and F. Phylogroups A and B1 were the most common phylogroups of E. coli obtained from healthy and sick-appearing broiler carcasses. The results of the study showed that phylogroups B2 and E were associated with the heart samples and phylogroup A was associated with respiratory tract samples, phylogroup B1 with not condemned carcass, and phylogroup D with liver samples.

Blood and milk polymorphonuclear leukocyte and monocyte/macrophage functions in naturally caprine arthritis encephalitis virus infection in dairy goats.

The exact influence of caprine arthritis encephalitis virus (CAEV) infection on blood and milk polymorphonuclear leukocytes (PMNLs) and monocyte/macrophages of goats remains unclear. Thus, the present study sought to explore the blood and milk PMNL and monocyte/macrophage functions in naturally CAEV-infected goats. The present study used 18 healthy Saanen goats that were segregated according to sera test outcomes into serologically CAEV negative (n=8; 14 halves) and positive (n=10; 14 halves) groups. All milk samples from mammary halves with milk bacteriologically positive outcomes, somatic cell count ≥2×106cellsmL-1, and abnormal secretions in the strip cup test were excluded. We evaluated the percentage of blood and milk PMNLs and monocyte/macrophages, the viability of PMNLs and monocyte/macrophages, the levels of intracellular reactive oxygen species (ROS) and the nonopsonized phagocytosis of Staphylococcus aureus and Escherichia coli by flow cytometry. In the present study, a higher percentage of milk macrophages (CD14+) and milk polymorphonuclear leukocytes undergoing late apoptosis or necrosis (Annexin-V+/Propidium iodide+) was observed in CAEV-infected goats; we did not find any further alterations in blood and milk PMNL and monocyte/macrophage functions. Thus, regarding our results, the goats naturally infected with CAEV did not reveal pronounced dysfunctions in blood and milk polymorphonuclear leukocytes and monocytes/macrophages.

Survey of pyrethroid, macrocyclic lactone and antibacterial residues in bulk milk tank from Minas Gerais State, Brazil / Resíduos de piretróides, lactonas macrocíclicas e antimicrobianos em amostras de leite de tanque no estado de Minas Gerais

A survey of veterinary drug residues in bulk milk tank from Minas Gerais State, Brazil, was carried out through a broad scope analysis. Here, 132 raw milk samples were collected at 45 dairy farms in Minas Gerais from August 2009 to February 2010, and analyzed for 42 analytes, comprising pyrethroids, macrocyclic lactones and antibacterials, using liquid chromatography coupled with mass spectrometry in tandem mode and gas chromatography with electron capture detection. Within all milk samples, at least one veterinary drug residue was identified in 40 milk samples (30.30%) by confirmatory tests, whereas 16 samples (12.12%) showed the presence of at least two residues. With regard to the Brazilian maximum residue levels, 11 milk samples (8.33%) were non-compliant according to Brazilian Legislation. The veterinary drugs detected in the non-compliant milk samples include penicillin V (one sample), abamectin (one sample) and cypermethrin (nine samples). Furthermore, the antibacterial screening methods failed to identify most of the positive samples that were detected by confirmatory tests, leading to a large discrepancy between the screening and confirmatory antimicrobial tests. Thus, the present study indicated that the veterinary drugs residues still represents a great concern for the milk production chain.(AU)

Avaliou-se a presença de 42 analitos, incluindo piretróides, lactonas macrocíclicas e antimicrobianos em 132 amostras de leite de tanque proveniente de 45 propriedades leiteiras localizadas no Estado de Minas Gerais. Para tal, utilizou-se a cromatografia líquida acoplada a espectrofotometria de massas tandem e cromatografia gasosa com detector com captura de elétrons. Dentre todas as amostras de leite, 40 (30,30%) amostras de leite de tanque apresentaram a presença de pelo menos um analito, enquanto 16 amostras (12,12%) de leite demonstraram a presença de pelo menos dois analitos. Considerando os limites estabelecidos pela legislação brasileira, 11 amostras de leite (8,33%) seriam consideradas como não conforme. Ademais, os testes de triagem para detecção de antimicrobianos no leite não conseguiram identificar a maioria das amostras positivas nos testes confirmatórios, levando a grande discrepância entre estes testes. Desta forma, os resultados do presente estudo indicam que os períodos de descarte do leite, especialmente para piretróides, não foram plenamente respeitados por todos os produtores de leite. Além disto, uma discrepância entre os resultados dos testes confirmatórios e os testes de triagem foi observada.(AU)

Characterization of virulence factors and phylogenetic group determination of Escherichia coli isolated from diarrheic and non-diarrheic calves from Brazil.

This study aimed to detect virulence factors, pathovars, and phylogenetic groups of Escherichia coli strains obtained from feces of calves with and without diarrhea up to 70 days old and to determine the association between occurrence of diarrhea, phylogenetic groups, and pathovars. Phylo-typing analysis of the 336 E. coli strains isolated from calves with Clermont method showed that 21 (6.25 %) belong to phylogroup A, 228 (67.85 %) to phylogroup B1, 2 (0.6 %) to phylogroup B2, 5 (1.49 %) to phylogroup C, 57 (16.96 %) to phylogroup E, and 3 (0.9 %) to phylogroup F. Phylogroup D was not identified and 20 strains (5.95 %) were assigned as "unknown." The distribution of phylogenetic groups among pathovars showed that NTEC belong to phylogroups B1 (17) and C (4); EPEC to phylogroups B1 (6) and E (8); STEC to phylogroups A (5), B1 (56), B2 (2), C (1), and E (15); EHEC to phylogroups B1 (95) and E (5); and ETEC to phylogroups A (3), B1 (7), and E (10). The EAST-1 strains were phylogroups A (13), B1 (47), E (19), and F (3); E. coli strains of "unknown" phylogroups belonged to pathovars EPEC (1), EHEC (2), STEC (7), and EAST-1 strains (6). ETEC was associated with diarrhea (P = 0.002). Our study did not find association between the phylogenetic background and occurrence of diarrhea (P = 0.164) but did find some relationship in phylogenetic group and pathovar. The study showed that EHEC and STEC are classified as phylogroup B1, EAST-1 phylogroup A, ETEC, and EPEC phylogroup E.

Epidemiological characterization and risk evaluation associated with the presence of Megninia spp. in posture farms / Caracterização epidemiológica e avaliação de risco associados à presença de Megninia spp. em granjas de postura

ABSTRACT: This study was accomplished with the aim to evaluate and to characterize the risks for the presence of Megninia spp. in poultry farms from Minas Gerais, using for that Correspondence Analysis (CA). A secondary database was used and adapted for this study, with information from 402 chicken houses of 42 laying hens commercial properties. Variables used to compose the correspondence model were selected through tests of Qui-square being considered the significant result of P≤0,05. A risk index was built to the presence of Megninia spp. starting from variables considered as risk factors for the presence of those ectoparasites. The CA graph produced for the epidemiological characterization of the presence of Megninia spp. presented an accumulated qui-square value of 58,55%. There was an association of the risk index with the age ranges of the poultry, among the presence Megninia spp. and the use of mineral oil and acaricide in the ration and; finally, an association between the ruddy ground-dove (Columbina talpacoti) presence and the absence of mites of the Megninia genus. The results supply knowledge on the epidemiology of Megninia spp. In addition, these information can contribute in the decision, reducing the risks of possible infestations in poultry farms and the negative effects of that parasitism.

RESUMO: Este estudo foi realizado com o objetivo de avaliar e caracterizar os riscos para a presença de Megninia spp. em galpões de granjas de postura de Minas Gerais, utilizando para isso Análise de Correspondência (AC). Foi utilizado um banco de dados secundário e adaptado para este estudo, com informações de 402 galpões de 42 propriedades comerciais de galinhas poedeiras. As variáveis utilizadas para compor o modelo de correspondência foram selecionadas através de testes de Qui-quadrado (P≤0,05). Foi construído um índice de risco para a presença de Megninia spp. a partir de variáveis consideradas como fatores de risco para a presença desses ectoparasitos. O gráfico da AC produzido para a caracterização epidemiológica da presença de Megninia spp. apresentou um valor de qui-quadrado acumulado de 58,55%. Observa-se que houve uma associação do índice de risco com a faixa de idades das aves, entre a presença Megninia spp. e a utilização de óleo mineral e de acaricida na ração e, por último, uma associação entre a presença de rolinhas (Columbina talpacoti) e a ausência de ácaros do gênero Megninia. Estes resultados fornecem conhecimento sobre a epidemiologia de Megninia spp. Além disso, estas informações poderão contribuir na tomada de decisão diminuindo os riscos de possíveis infestações nos plantéis e dos efeitos negativos decorrentes por esse parasitismo.

Phylogenetic Group Determination of Escherichia coli Isolated from Animals Samples.

This study analyzes the occurrence and distribution of phylogenetic groups of 391 strains of Escherichia coli isolated from poultry, cattle, and water buffalo. The frequency of the phylogroups was A = 19%, B1 = 57%, B2 = 2.3%, C = 4.6%, D = 2.8%, E = 11%, and F = 3.3%. Phylogroups A (P < 0.001) and F (P = 0.018) were associated with E. coli strains isolated from poultry, phylogroups B1 (P < 0.001) and E (P = 0.002) were associated with E. coli isolated from cattle, and phylogroups B2 (P = 0.003) and D (P = 0.017) were associated with E. coli isolated from water buffalo. This report demonstrated that some phylogroups are associated with the host analyzed and the results provide knowledge of the phylogenetic composition of E. coli from domestic animals.

The neutrophil function and lymphocyte profile of milk from bovine mammary glands infected with Streptococcus dysgalactiae.

Streptococcus dysgalactiae is a bacterium that accounts for a notable proportion of both clinical and subclinical intramammary infections (IMIs). Thus, the present study explores the function of milk neutrophils and the lymphocyte profile in mammary glands naturally infected with Streptococcus dysgalactiae. Here, we used 32 culture-negative control quarters from eight clinically healthy dairy cows with low somatic cell counts and 13 S. dysgalactiae-infected quarters from six dairy cows. Using flow cytometry, we evaluated the percentage of milk monocytes/macrophages and neutrophils, expression of CD62L, CD11b and CD44 by milk neutrophils, the levels of intracellular reactive oxygen species (ROS) production and phagocytosis of Staphylococcus aureus by milk neutrophils, and neutrophil viability. Furthermore, the percentages of B cell (CD21(+)) and T lymphocyte subsets (CD3(+)/CD4(+)/CD8(-); CD3(+)/CD8(+)/CD4(-); and CD3(+)/CD8(-)/CD4(-)), and the expression of CD25 by T milk lymphocytes (CD3(+)) and T CD4(+) milk cells were also assessed by flow cytometry using monoclonal antibodies. The present study showed a higher SCC and percentage of milk neutrophils, and a decrease in the percentage of milk monocytes/macrophages from S. dysgalactiae-infected quarters when compared to uninfected ones. We also observed a higher expression of CD11b by milk neutrophils and a tendency toward a decrease in neutrophil apoptosis rate in S. dysgalactiae-infected quarters. In addition, the S. dysgalactiae-infected quarters had higher percentages of milk T cells (CD3(+)) and their subset CD3(+)CD8(+)CD4(-) cells. Overall, the present study provided new insights into S. dysgalactiae IMIs, including distinct lymphocyte profiles, and a tendency toward an inhibition of apoptosis in milk neutrophils.

Effects of bovine leukemia virus infection on milk neutrophil function and the milk lymphocyte profile.

The effects of bovine leukemia virus (BLV) on the immune response have been extensively investigated; however, its effects on mammary gland immunity are only speculative. Although BLV has a tropism for B cells, it can affect both adaptive and innate immunities because these systems share many effector mechanisms. This scenario is the basis of this investigation of the effects of BLV on mammary gland immunity, which is largely dependent upon neutrophilic functions. Thus, the present study sought to examine neutrophilic functions and the lymphocyte profile in the milk of naturally BLV-infected cows. The viability of the milk neutrophils and the percentage of milk neutrophils that produced reactive oxygen species (ROS) or phagocytosed Staphylococcus aureus were similar between BLV-infected and BLV-uninfected dairy cows. Furthermore, the expression of CD62L and CD11b by the milk neutrophils and the percentage of milk neutrophils (CH138+ cells) that were obtained from the udder quarters of the BLV-infected cows were not altered. Conversely, the median fluorescence intensity (MFI) representing intracellular ROS production and the phagocytosis of S. aureus, the expression of CD44 by the milk neutrophils and the percentage of apoptotic B cells were lower in the milk cells from BLV-infected dairy cows, particularly those from animals with persistent lymphocytosis (PL). The lymphocyte subsets were not different among the groups, with the exception of the percentage of CD5-/CD11b- B cells, which was higher in the milk cells from BLV-infected cows, particularly those with PL. Thus, the present study provides novel insight into the implications of BLV infection for mammary gland immunity.

Influence of race and crossbreeding on casein micelles size.

Casein (CN) micelles are colloidal aggregates of protein dispersed in milk, the importance of which in the dairy industry is related to functionality and yield in dairy products. The objective of this work was to investigate the correlation of milk CN micelles diameter from Holstein and Zebu crossbreds with milk composition (protein, fat, lactose, total and nonfat solids and milk urea nitrogen), somatic cell count (SCC), age, lactation stage and production. Average casein micelles diameters of milk samples obtained from 200 cows were measured using photon correlation spectroscopy and multiple regression analysis was used to find relationship between variables. CN micelle diameter, SCC and nonfat solids were different between animals with different Holstein crossbreed ratios, which suggests influence of genetic factors, mammary gland health and milk composition. Overall, results indicate the potential use of CN micelle diameter as a tool to select animals to produce milk more suitable to cheese production.