Perfil microbiológico da mastite bovina no Agreste e Brejo Paraibano

Bovine mastitis is the most common disease in dairy herds worldwide, affecting producers, the dairy industry and milk quality, in addition to having a negative impact on public health. This study aimed to follow the microbiological profile of the causative agents of bovine mastitis in dairy farms in the Agreste and Brejo microregion of Paraiba from August 2018 to December 2019. A total of 421 milk samples were collected from 987 mammary quarters of 250 cows diagnosed with clinical (CM) and subclinical  mastitis (MSC) . MSC had a higher frequency of cases 405 (41.03 %) than MC 51 (5.16%). In the microbiological analyses, 16.63% of the samples collected from animals with MC or MSC were found to be negative. The most isolated genera in microbiological growth amostras were Staphylococcus spp. (26,36 %), Corynebacterium spp. (22,32 %), and Streptococcus spp. (12,59 %), while environmental agents showed sporadic cases in their isolation. In the evaluation of the CMT dynamics, it was found that samples with positive CMT results with negative microbiological exam result in M1, tended to have a significant reduction in CMT scores in M2. The findings highlight the importance of conducting health education in the region's milking process, as well as reinforcing the importance of using CMT in a systematic and criterion-based manner.A mastite bovina é considerada a doença de maior prevalência nos rebanhos leiteiros no mundo, impactando produtores, indústria de laticínios e influenciando negativamente a qualidade do leite, além do impacto na saúde pública. Nesse contexto, objetivou-se traçar o perfil microbiológico dos agentes causadores da mastite bovina em propriedades leiteiras da microrregião do Agreste e Brejo paraibano entre o período de agosto de 2018 a dezembro de 2019 e avaliar o comportamento da dinâmica do California mastitis test (CMT) em diferentes perfis microbiológicos do leite. Foram analisadas um total de 421 amostras de leite provenientes de 920 quartos mamários de 230 vacas que foram submetidas ao diagnóstico da mastite clínica (MC) e subclínica (MSC). Observou-se uma expressiva ocorrência de casos de MSC (n = 370, 40,22%) e de MC (n = 51, 5,54%). As amostras de quartos mamários com MC ou MSC resultou em 16,63% de ausência de crescimento no exame microbiológico. Das amostras com crescimento microbiológico, Staphylococcus spp. (26,36%), Corynebacterium spp. (22,32%) e Streptococcus spp. (12,59%) foram os microrganismos mais frequentemente isolados, já os agentes etiológicos ambientais foram esporadicamente isolados. Na avaliação da dinâmica do CMT observou-se que as amostras com resultado positivo no CMT e negativos no exame microbiológico apresentaram tendência a uma redução expressiva dos escores de CMT no momento da coleta subsequente (após 30 dias). Os resultados demonstram a necessidade de realizar educação no processo de ordenha na região, bem como, reforçar a importância do uso do CMT e do exame microbiológico de forma criteriosa no controle da mastite bovina

Brief Research Report: Expression of PD-1 and CTLA-4 in T Lymphocytes and Their Relationship With the Periparturient Period and the Endometrial Cytology of Dairy Cows During the Postpartum Period

The present study sought to evaluate the expression of PD-1 and CTLA-4 in blood T lymphocytes during the periparturient period and their relationship with uterine health in dairy cows, as determined by endometrial cytology and serum concentrations of β-hydroxybutyrate (BHB) and non-esterified fatty acids (NEFAs), which are indicators of a negative energy balance. The second objective of this study was to investigate whether the expression of PD-1 and CTLA-4 in T lymphocytes is associated with the serum acute phase-protein haptoglobin concentration during the periparturient period. To address these objectives, 26 clinically healthy dairy cows were used. Peripheral blood was collected 14 days prepartum (T-14), at calving (T0), and 30 days postpartum (T30) to measure the expression of PD-1 and CTLA-4 in blood T lymphocytes by flow cytometry. In addition, we collected blood at T0, 10 days after parturition (T10), and T30 to obtain serum and determine the serum concentrations of NEFA, BHB, and Hp. Endometrial cytology was performed at T10, 20 days after parturition (T20), and T30. In the present study, we observed higher expression of CTLA-4 and PD-1 in T lymphocytes at parturition and in the prepartum period, which could indicate a relationship between these immune checkpoints and immunological tolerance during gestation in dairy cattle. In addition, a negative association between the expression of these immune checkpoints prepartum or at parturition and endometrial cytology at T20 and T30 was observed, indicating the negative implications of these immune response regulators in susceptibility to infections. This finding was further corroborated by the relationship between the serum concentration of haptoglobin and the expression of CTLA-4 and PD-1 by T lymphocytes. However, we did not observe a relationship between the indicators of negative energy balance, evaluated by the serum concentrations of BHB and NEFA, and the expression of the immune checkpoint markers studied. Thus, our findings represent an initial step that paves the way for the development of new therapeutic alternatives directed by the host with the objective of increasing the resistance of dairy cattle to infections in this critical period of life

Efeito de infusões intravenosas e intramamárias de lipopolissacarídeo (LPS) sobre parâmetros inflamatórios, reprodutivos e metabólicos em novilhas e vacas da raça holandesa

Objetivou-se avaliar os efeitos da resposta inflamatória na expressão gênica endometrial e no metabolismo energético em novilhas e vacas. Inicialmente, foi estabelecido um protocolo para indução de um perfil pró-inflamatório sistêmico, com base em múltiplas infusões de LPS. Posteriormente, 22 novilhas foram sincronizadas, inseminadas (D0) e divididas em grupo controle (sol. salina) e LPS, iniciando as infusões no D2. O grupo LPS recebeu infusões intravenosas (IV) de solução salina + LPS (0,1; 0,25; 0,5; 0,75; 1,0; 1,25 mg/Kg). Além da avaliação da temperatura retal, foram analisadas as concentrações plasmáticas de TNF-α, haptoglobina (Hp), progesterona, AGNE, e BHB, além das avaliações leucométricas. No D15, foram realizadas as biópsias endometriais, e posterior análise por qRT-PCR de genes relacionados ao adesão celular, sistema imunológico e desenvolvimento embrionário. As infusões de LPS estabeleceram um perfil próinflamatório caracterizado pelo aumento da temperatura (P<0,05), dos níveis de TNF-α (P=0,05) e de Hp (P<0,01), além de alteração na contagem diferencial de leucócitos. Não foi observada alteração nos parâmetros metabólicos. Em relação ao grupo controle, o grupo LPS apresentou menor expressão de IDO (P=0,04; fold=0,48) e PTX3 (P=0,01; fold=0,38), e uma expressão elevada de MX1 (P=0,02; fold=2,85). Em um segundo trabalho, para avaliar a influência da resposta inflamatória sobre o metabolismo energético de vacas em lactação, 20 vacas foram divididas em grupos controle (10 mL sol. Salina) e LPS (25 μg LPS/10 mL sol. salina). Foi avaliada a contagem de células somáticas (CCS), a produção leiteira, temperatura reticular, além dos níveis sorológicos de Hp, TNF-α, AGNE e BHB. No grupo LPS foi observado o aumento da CCS (P<0,0001), queda da produção (P<0,05), além do aumento da temperatura reticular (P<0,005) e aumento da concentração da Hp (P<0,0001). Verificou-se aumento de AGNE no dia nove e de BHB nos dias nove (P<0,005) e 10 (P=0,05). Com a realização dos presentes trabalhaos conclui-se inicialmente o protocolo utilizado no primeiro trabalho induziu um marcante perfil próinflamatório, porém, com limitado efeito na expressão do gene relacionados ao sistema imunitário, o que sugere uma possível explicação para a sub-fertilidade relacionadas a alterações fisiopatológicas em vacas leiteiras. Adicionalemnte, considerando a resposta inflamatória induzida pelo LPS intramamário, foi possível observar evidências da relação da resposta inflamatória com o metabolismo energético de vacas em lactação, e reafirmar a participação de mediadores inflamatórios na fisiopatogenia de enfermidades metabólicas em vacas de leite.The aim of the study was to evaluate the effects of the inflammatory response over endometrial gene expression and energy metabolism in heifers and cows. Initially, a protocol to induce a systemic pro-inflammatory profile was established based on multiple LPS infusions. Subsequently, 22 heifers were synchronized, inseminated (D0) and divided into control group (Saline sol.) and LPS, infusions started on D2. LPS group received intravenous infusions (IV) with saline solution + LPS (0.1; 0.25; 0.5; 0.75; 1.0; 1.25 mg / kg). Were analyzed the rectal temperature, plasma concentrations of TNF-α, haptoglobin (Hp), progesterone, AGNE and BHB, beside leucometrics analysis. On D15, the endometrial biopsies were performed, and subsequent analysis by qRT-PCR of genes related to cell adhesion, immune system and embryonic development. LPS infusions established a pro-inflammatory profile characterized by increase of temperature (P<0.05), TNF-α (P=0.05) and Hp (P<0.01), and effects on the differential leukocyte count. There was no change in metabolic parameters. The LPS group showed lower expression of IDO (P=0.04; fold=0.48) and PTX3 (P=0.01; fold=0.38), and high expression of MX1 (P=0.02; fold=2.85). Was concluded that the induced pro-inflammatory profile showed a strong but limited effect on gene expression related to immune system, suggesting a possible explanation for sub-fertility related pathophysiological changes in dairy cows. In the second study, to evaluate the influence of the inflammatory response on the energy metabolism of dairy cows, 20 cows were divided into control group (10 mL saline sol.) and LPS (25 ug LPS/10ml saline sol.). Was evaluated the somatic cell count, milk production, reticular temperature, as well the serum levels of Hp, TNF-α, AGNE and BHB. In LPS group was observed increased SCC (P<0.0001), decrease of production (P<0.05), and increased reticular temperature (P<0.005), as well an increase of Hp concentration (P<0,0001). Was observed increase on AGNE on day nine and BHB on day nine (P<0.005) and ten (P=0,05). Was concluded that the induced pro-inflammatory profile showed a strong but limited effect on gene expression related to immune system, suggesting a possible explanation for sub-fertility related pathophysiological changes in dairy cows. Aditonally, considering the inflammatory response induced by LPS intramammary infusions, were observed evidences of the relationship of the inflammatory response with the energy metabolism of dairy cows, and reaffirmed the participation of inflammatory mediators in the pathogenesis of metabolic diseases in dairy cows.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPE

Avaliação da leucometria na identificação da leucose enzoótica dos bovinos em rebanhos do estado de Pernambuco

A Leucose Enzoótica dos Bovinos (LEB) é uma ameaça à saúde dos rebanhos bovinos do estado de Pernambuco e o aperfeiçoamento no uso de ferramentas epidemiológicas para identificação e eliminação de focos desta doença demanda atenção. O objetivo com a realização deste estudo foi avaliar a leucometria como um recurso auxiliar para a identificação de focos da LEB e saneamento dos rebanhos, a partir de ensaios realizados para estabelecer a prevalência dessa insidiosa retrovirose e da Tuberculose Bovina (TB) em rebanhos criados em vários municípios do estado. Foram submetidas ao sorodiagnóstico para LEB amostras de 1.000 bovinos procedentes de 33 rebanhos, sendo 920 deles submetidos previamente ao teste da tuberculina (TCC). Em aproximadamente 70% (694/1000) dos bovinos foram realizadas análises leucométricas, sendo desconsiderados das mesmas os animais com resultados inconclusivos aos dois testes diagnósticos e os positivos ao teste da tuberculina. Desta forma, 530 amostras foram distribuídas em dois grupos experimentais, em função dos resultados à IDGA: GI = leucogramas de 379 bovinos soronegativos; e GII = leucogramas de 151 bovinos soropositivos. As prevalências da LEB e TB foram 28% (282/1000) e 11% (99/920), respectivamente. Globalmente, os valores médios dos leucócitos foram: totais = 12,0 ± 4,7 e linfócitos = 8,1 ± 5,4 (x103/mm3). Considerando os grupos experimentais, os valores médios dos leucócitos do GI (totais = 11,5 ± 3,8 e linfócitos = 7,6 ± 5,1 x103/mm3) foram significativamente menores (p 15,0 e linfócitos > 12,7 x103/mm3) foram considerados suspeitos, pois não apresentaram diferença leucométrica significativa com os do GII (soropositivos) portadores de LL. Conclui-se que, com a demonstração da interferência do VLB no leucograma dos bovinos examinados e com a identificação de bovinos suspeitos não detectados pela IDGA, a leucometria se presta como recurso auxiliar para a identificação de focos da LEB e saneamento dos rebanhos.The Enzootic Bovine Leukosis (EBL) is a threat to the health of cattle herds in the state of Pernambuco and improvement in the use of epidemiological tools to identify and eliminate outbreaks of the disease demands attention. The aim of this study was to evaluate the leukocyte count as an aid in the identification of outbreaks of EBL and sanitation of livestock, from tests performed to establish the prevalence of this insidious retrovirus and Bovine Tuberculosis (BT) in cattle raised in several cities in the state. Were submitted for the EBL´s serodiagnosis 1000 bovine samples originating from 33 herds, 920 of them being previously submitted to the tuberculin test. Approximately 70% (694/1000) of the bovine leukocyte counts were analyzed, and disregarded animals with inconclusive results to two diagnostic tests and the positive to the tuberculin test. Thus, 530 samples were divided into two experimental groups, according to the results of AGID: GI = white blood cell count of 379 seronegative cattle, and IGI = white blood cell count of 151 seropositive cattle. The prevalence of EBL and BT were 28% (282/1000) and 11% (99/920), respectively. Overall, the mean values of leukocytes were: total = 12.0 ± 4.7 and lymphocytes = 8.1 ± 5.4 (x103/mm3). Considering the experimental groups, the mean values of leukocytes in GI (total = 11.5 ± 3.8 and = 7.6 ± 5.1 lymphocytes x103/mm3) were significantly lower (p 15.0 and lymphocytes> 12.7 x103/mm3) were considered suspect because leukocyte counts did not differ significantly with those of GII (seropositive) with LL. It is concluded that with the VLB demonstration of interference in leukocyte counts of cattle examined and the identification of suspect cattle not detected by AGID, the leukocyte count lends itself as an auxiliary to the identification of outbreaks of LEB and sanitation of livest

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

Neotropical freshwater fisheries : A dataset of occurrence and abundance of freshwater fishes in the Neotropics

The Neotropical region hosts 4225 freshwater fish species, ranking first among the world's most diverse regions for freshwater fishes. Our NEOTROPICAL FRESHWATER FISHES data set is the first to produce a large-scale Neotropical freshwater fish inventory, covering the entire Neotropical region from Mexico and the Caribbean in the north to the southern limits in Argentina, Paraguay, Chile, and Uruguay. We compiled 185,787 distribution records, with unique georeferenced coordinates, for the 4225 species, represented by occurrence and abundance data. The number of species for the most numerous orders are as follows: Characiformes (1289), Siluriformes (1384), Cichliformes (354), Cyprinodontiformes (245), and Gymnotiformes (135). The most recorded species was the characid Astyanax fasciatus (4696 records). We registered 116,802 distribution records for native species, compared to 1802 distribution records for nonnative species. The main aim of the NEOTROPICAL FRESHWATER FISHES data set was to make these occurrence and abundance data accessible for international researchers to develop ecological and macroecological studies, from local to regional scales, with focal fish species, families, or orders. We anticipate that the NEOTROPICAL FRESHWATER FISHES data set will be valuable for studies on a wide range of ecological processes, such as trophic cascades, fishery pressure, the effects of habitat loss and fragmentation, and the impacts of species invasion and climate change. There are no copyright restrictions on the data, and please cite this data paper when using the data in publications

NEOTROPICAL ALIEN MAMMALS: a data set of occurrence and abundance of alien mammals in the Neotropics

Biological invasion is one of the main threats to native biodiversity. For a species to become invasive, it must be voluntarily or involuntarily introduced by humans into a nonnative habitat. Mammals were among first taxa to be introduced worldwide for game, meat, and labor, yet the number of species introduced in the Neotropics remains unknown. In this data set, we make available occurrence and abundance data on mammal species that (1) transposed a geographical barrier and (2) were voluntarily or involuntarily introduced by humans into the Neotropics. Our data set is composed of 73,738 historical and current georeferenced records on alien mammal species of which around 96% correspond to occurrence data on 77 species belonging to eight orders and 26 families. Data cover 26 continental countries in the Neotropics, ranging from Mexico and its frontier regions (southern Florida and coastal-central Florida in the southeast United States) to Argentina, Paraguay, Chile, and Uruguay, and the 13 countries of Caribbean islands. Our data set also includes neotropical species (e.g., Callithrix sp., Myocastor coypus, Nasua nasua) considered alien in particular areas of Neotropics. The most numerous species in terms of records are from Bos sp. (n = 37,782), Sus scrofa (n = 6,730), and Canis familiaris (n = 10,084); 17 species were represented by only one record (e.g., Syncerus caffer, Cervus timorensis, Cervus unicolor, Canis latrans). Primates have the highest number of species in the data set (n = 20 species), partly because of uncertainties regarding taxonomic identification of the genera Callithrix, which includes the species Callithrix aurita, Callithrix flaviceps, Callithrix geoffroyi, Callithrix jacchus, Callithrix kuhlii, Callithrix penicillata, and their hybrids. This unique data set will be a valuable source of information on invasion risk assessments, biodiversity redistribution and conservation-related research. There are no copyright restrictions. Please cite this data paper when using the data in publications. We also request that researchers and teachers inform us on how they are using the data