Virulence-encoding genes related to extraintestinal pathogenic E. coli and multidrug resistant pattern of strains isolated from neonatal calves with different severity scores of umbilical infections.

Umbilical infections in calves comprise a major cause of neonatal mortality and have been related to a variety of microorganisms. E. coli is an opportunistic enteropathogen characterized by a diversity of virulence factors (VF). Nonetheless, the gene profiles that encode VF associated with umbilical infections in calves and their effect on the clinical severity remains unclear. In this scenario, microbial identification (with an emphasis on E. coli), was carried out among 150 neonatal calves (≤30 days of age) with umbilical infections, where the omphalopathies were clinically scored as mild, moderate, or severe. Also, a panel of 16 virulence-encoding genes related to extraintestinal pathogenic E. coli (ExPEC) were investigated, i.e., fimbriae/adhesins (sfa/focDEa, papA, papC, afaBC), toxins (hlyA, sat, cnf1, cdt), siderophores (iroN, irp2, iucD, ireA), invasins (ibeA), and serum resistance (ompT, traT, kpsMT II). Bacteria and yeasts isolates were identified using mass spectrometry. Bacteria, yeasts, and fungi were isolated in 94.7% (142/150) of neonatal calves sampled. E. coli was the agent most frequently isolated (59/150 = 39.3%), in pure culture (27/59 = 45.8%) and combined infections (32/59 = 54.2%), although a great variety (n = 83) of other species of microorganisms were identified. Clinical severity scores of 1, 2, and 3 were observed in 32.2% (19/59), 23.7% (14/59), and 44.1% (26/59) of E. coli infections, respectively. The ExPEC genes detected were related to serum resistance (traT, 42/59 = 72.2%; ompT, 35/59 = 59.3%, kpsMTII, 10/59 = 17%), invasins (ibeA, 11/59 = 18.6%), siderophores (iucD, 9/59 = 15.3%; iroN, 8/59 = 13.6%), and adhesins/fimbriae (papA, 8/59 = 13.6%; papC, 15/59 = 9.6%). The presence of each virulence gene was not associated with the case's clinical score. Among all isolates, 89.8% (53/59) showed in vitro resistance to sulfamethoxazole/trimethoprim and 59.3% to ampicillin (35/59), while 94.1% (55/59) revealed a multidrug resistant profile. Great complexity of bacteria, yeast, and fungi species was identified, reinforcing the umbilical infections of neonatal calves as a polymicrobial disorder. The high occurrence of E. coli (39.3%) highlights the role of this pathogen in the etiology of umbilical infections in calves. Furthermore, a panel of ExPEC genes was investigated for the first time among calves that were clinically scored for case severity. The high prevalence of traT and ompT indicates that these serum resistance-related genes could be used as biomarkers for further investigations of ExPEC isolates from umbilical infections. Our results contribute to the etiological investigation, clinical severity scoring, antimicrobial resistance pattern, and virulence-related to ExPEC genes involved in umbilical infections of neonatal calves.

Safflower seed supplementation in lamb feed: effects upon fatty acid profile and quality of meat patty formulations.

The aims of this study were to determine the fatty acid profile of meat from lambs fed with different levels of safflower seed (0%, 7.5%, and 15%) and, also, to compare the characteristics of the meat patties prepared from this lamb meat (LMP) with beef meat patties (BMP). The safflower seed-supplemented diet did not change the contents of polyunsaturated and unsaturated fatty acids, except for C22:1. All meat patty formulations were considered safe for consumption. The values of cooking yield, shrinkage, water absorption index, luminosity (L*), and redness (a*) were similar for the LMP and BMP tested. While the safflower seed-supplemented diet did not alter the moisture, ash, and protein levels of LMP, the lipid content was lower than that in BMP. The incorporation of 15% safflower seed into lamb feed contributed to promoting better sensory attributes of the meat patties. Most of the physicochemical properties evaluated were similar among LMP and BMP. However, to improve the sensory properties of the product, dietary supplementation with 15% safflower seed is recommended.

Occurrence of brucellosis, leptospirosis and neosporosis in cows with retained placenta in Southwest Paraná, Brazil / Ocorrência de brucelose, leptospirose e neosporose em vacas com retenção de placenta no Sudoeste do Paraná, Brasil

The aim of this study was to measure the occurrence of brucellosis, leptospirosis and neosporosis in cows from 25 family farms milk, with semi-intensive grazing system, located in the Southwest region of Paraná, Brazil. Eighty-four cows with retained placenta, between July 2013 to July 2014, diagnosed by the presence of fetal membranes in the uterus up to 12 hours after partum, decrease of appetite and of milk production, as well increase of rectal temperature were included into the study. The animals were submitted to blood collection for evaluating the seropositivity for brucellosis, leptospirosis and neosporosis. The blood was collected at 15 to 45 days postpartum, avoiding false negative results due to immune deficiency observed in the transition period. After collection, the blood was centrifuged, the serum was packed in three aliquots, identified and subsequently frozen. Serology was performed to diagnose brucellosis (technique of buffered acidified antigen), leptospirosis (through the microscopic agglutination test - MAT) and neosporosis (indirect immunofluorescence test). Animals were considered positive when presenting titer >1:100 (leptospirosis) and 1:200 (neosporosis). No animal with placenta retention presented Brucella abortus bacteria; 39% (33/84) were reactive to one or more Leptospira serovars, 15% (13/84) were positive to Neospora caninum, and 4% (4/84) had both diagnosis, being reactive for leptospirosis and neosporosis. In conclusion, data from this experiment inspires greater attention to leptospirosis and neosporosis in dairy cattle presenting placenta retention in southwest Paraná.(AU)

Objetivou-se com esse estudo avaliar a ocorrência de brucelose, leptospirose e neosporose em fêmeas bovinas de 25 propriedades leiteiras de agricultura familiar, mantidas em sistema de pastejo semi-intensivo, localizadas na Região Sudoeste do Paraná, Brasil. Para tanto, incluiu-se nesse estudo, 84 vacas diagnosticadas com retenção de placenta (RP) pela presença parcial ou total dos anexos fetais no útero após 12 horas do parto, diminuição do apetite e da produção de leite, como também aumento da temperatura retal. Os animais foram submetidos à coleta de sangue 15 a 45 dias pós-parto, a fim de evitar resultado falso negativo devido à imunodeficiência observada no período de transição. Após coleta, o sangue foi centrifugado, o soro aliquotado em três amostras, identificado e congelado. Para o diagnóstico sorológico de Brucelose utilizou-se o exame sorológico do antígeno acidificado tamponado, de leptospirose a técnica de soroaglutinação microscópica, e neosporose pela técnica de imunofluorescência indireta. Os animais foram diagnosticados positivos quando apresentaram título igual ou superior à 1:100 (leptospirose) e 1:200 (neosporose). Em nenhum dos 84 animais com RP foi detectada a presença da bactéria Brucella abortus. Desses, 39,28% (33/84) foram reativos para um ou mais sorovares de Leptospira, 15,47% (13/84) foram positivos para Neospora caninum e 4,76% (4/84) foram reativos para sorovares de Leptospira sp. e N. caninum, concomitantemente. Em conclusão, os dados deste estudo inspiram maior atenção à leptospirose e neosporose em bovinos leiteiros que apresentam retenção de placenta no sudoeste do Paraná.(AU)

Colheita de sêmen por eletroejaculação em cutia-parda (Dasyprocta azarae) / Semen collection by electroejaculation in Azara´s agouti (Dasyprocta azarae)

As cutias são animais que vivem nas Américas Central e do Sul, com potencial zootécnico e que despertam crescente interesse científico. O objetivo deste estudo foi estabelecer um padrão para colheita de sêmen por eletroejaculação em cutia-parda (Dasyprocta azarae). Foram utilizados machos adultos (n=4) anestesiados e submetidos a quatro séries de 20 estímulos eletroejaculatórios, com 3 segundos de duração cada, com uma intensidade de 2, 4, 6 e 8 volts, com dois minutos de intervalo entre cada série. A colheita de sêmen foi obtida em todos os animais, com ereção parcial com 2 volts, ereção evidente com 4 volts, ejaculação com 6 volts e exacerbação da glande e exteriorização das espículas penianas com 8 volts, observando-se espermatozoides em 100% das amostras. A técnica descrita para colheita de sêmen em cutia-parda foi eficaz.

Agoutis are animals that live in South and Central America, present rearing potential and call increasing scientific interest. The aim of this paper was to establish a model for semen collection by electroejaculation in Azara´s agouti (Dasyprocta azarae). Anesthetized adult males (n=4) were submitted to four series of 20 electroejaculatory stimuli, during 3 seconds each and with 2, 4, 6 and 8 volts, with a 2 minutes rest among series. Semen collection was accomplished in every animal, with partial penis erection at 2 volts, makeable erection at 4 volts, ejaculation at 6 volts, glans and penis spines exposition at 8 volts, and sperm collected in all samples. The described technique for Azara's agouti semen collection was efficient.

Características reprodutivas de veado-bororó-do-sul ou veado-mão-curta (Mazama nana) / Reproductive characteristics of Brazilian dwarf brocket deer (Mazama nana)

Dos cervídeos brasileiros, a espécie Mazama nana é a menos conhecida. Os parâmetros reprodutivos para os machos ainda são desconhecidos, mas parece que não apresentam sazonalidade reprodutiva. Neste trabalho foram utilizados nove machos de Mazama nana em idade reprodutiva, mantidos em cativeiro. Foram avaliados quanto ao peso corporal, altura de cernelha, comprimento crânio-caudal, situação dos chifres, volume e consistência testicular. O sêmen foi colhido por eletroejaculação e submetido a análises de motilidade, vigor e morfologia espermática. As correlações entre idade, peso, comprimento crânio-caudal, altura de cernelha, volume testicular e características do ejaculado (volume, motilidade, vigor e concentração do sêmen) foram avaliadas pelo procedimento Corr., do SAS®. As médias ± desvio padrão observados para peso (kg), comprimento crânio-caudal (cm) e altura de cernelha (cm) foram: 15,72±1,98, 74,9±3,05 e 48,5±2,06, respectivamente. Em relação aos parâmetros reprodutivos primários foram observados: volume do ejaculado (91,46±68,24µl); motilidade (70±8,16 por cento); vigor (3,0±0,67); concentração espermática (1536x10(6) ±351x10(6) espermatozóides por ml). Em relação à morfologia espermática, foi observada uma alta porcentagem de células anormais (40,90 por cento), sendo predominante os defeitos de cauda (25,95 por cento).

Mazama nana is the least known of Brazilian deer species. The male reproductive parameters are still unknown, but apparently they did not show reproductive seasonality. In this work were used nine males of M. nana in reproductive age, kept in captivity. They were assessed for weight, height of wither, crown-rump length, the situation of horns, testicular volume and consistency. The sperm was collected by electro-ejaculation and subjected to analyses of motility, vigor and morphology. The correlation between age, weight, height of wither, crown-rump length, testicular volume and characteristics of the ejaculate (volume, motility, vigor and sperm concentration) were evaluated by the procedure Corr., of SAS®. The mean ± standard deviation observed for weight (kg), crown-rump length (cm) and height of wither (cm) were: 15.72±1.98, 74.9±3.05, and 48.5±2.06 respectively. For primary reproductive parameters were observed: volume of ejaculate (91.46±68.24ìl); motility (70±8.16 percent); vigor (3.0±0.67); sperm concentration (1536x10(6) ±351x10(6) spermatozoa per ml). For sperm morphology was found a high percentage of abnormal cells (40.90 percent), with predominant defects in the tail (25.95 percent).