Use of new recombinant proteins for ovarian stimulation in ruminants

Currently, gonadotropin products (follicle stimulating hormone, FSH, and luteinizing hormone, LH) used in animal reproduction are produced by extraction and purification from abattoir-derived pituitary glands. This method, relying on animal-derived materials, carries the potential risk of hormone contamination and pathogen transmission. Additionally, chorionic gonadotropins are extracted from the blood of pregnant mares (equine chorionic gonadotropin; eCG) or the urine of pregnant women (human chorionic gonadotropin; hCG). However, recent advancements have introduced recombinant gonadotropins for assisted animal reproduction therapies. The traditional use of FSH for superovulation has limitations, including labor requirements and variability in superovulation response, affecting the success of in vivo (SOV) and in vitro (OPU/IVEP) embryo production. FSH treatment for superstimulation before OPU can promote the growth of a homogenous follicular population and the recovery of competent oocytes suitable for IVEP procedures. At present, a single injection of a preparation of long-acting bovine recombinant FSH (rFSH) produced similar superovulation responses resulting in the production of good-quality in vivo and in vitro embryos. Furthermore, the treatment with eCG at FTAI protocol has demonstrated its efficacy in promoting follicular growth, ovulation, and P/AI, mainly in heifers and anestrous cows. Currently, treatment with recombinant glycoproteins with eCG-like activity (r-eCG) have shown promising results in increasing follicular growth, ovulation, and P/AI in cows submitted to P4/E2 -based protocols. Bovine somatotropin (bST) is a naturally occurring hormone found in cows. Recombinant bovine somatotropin (rbST), produced through genetic engineering techniques, has shown potential in enhancing reproductive outcomes in ruminants. Treatment with rbST has been found to improve P/IA, increase donor embryo production, and enhance P/ET in recipients. The use of recombinant hormones allows to produce non-animal-derived products, offering several advantages in assisted reproductive technologies for ruminants. This advancement opens up new possibilities for improving reproductive efficiency and success rates in the field of animal reproduction.(AU)

Real-time polymerase chain reaction for detection and enumeration of Staphylococcus aureus and Streptococccus agalactiae using different milk samplings

The objective of the present study was to evaluate the qPCR for detection and enumeration of Staphylococcus aureus and Streptococcus agalactiae using different milk samplings in comparison to the conventional microbiology. Four dairy herds with a history of subclinical mastitis caused by S. aureus and S. agalactiae were selected. Sampling approach included milk samples from bulk tank (BT), cow level (composite samples, CO), and mammary quarter level (MQ) from 785 lactating cows. Three consecutive monthly milk samplings were carried out, totaling 3347 MQ milk samples, 912 CO, and 12 from BT. All collected milk samples were subjected to conventional microbiology and qPCR for detection and enumeration of S. aureus and S. agalactiae. The qPCR showed 71.5% of diagnostic sensitivity for S. aureus isolated from MQ milk samples, 71.8% for CO, and 50% for BT milk samples compared with conventional microbiology methodology. Taken together, the diagnostic sensitivity for S. agalactiae isolated from MQ milk samples was 90.2, 87.7 for CO, and 90.9% for BT milk samples. In general, the qPCR methodology enabled the detection of S. aureus and S. agalactiae, regardless of the type of milk sampling. The direct use of milk samples to estimate the counting of S. aureus by qPCR demonstrated lower sensitivity than the counting of S. agalactiae, which can be explained by the pathogen infection dynamics and differences in milk sample type.

Seroprevalence and risk factors associated with leukosis in cattle from Villlavicencio, Colombia / Soroprevalência e fatores de risco associados à leucose em bovinos de Villlavicencio, Colômbia

Bovine leukosis is caused by an oncogenic virus of the genus Deltaretrovirus, causing losses associated with decreased production indicators and restrictions on exports of cattle and cattle products. The disease has a prolonged incubation period of between 1-5 years and the antibodies can be detected 2-3 weeks post infection. The disease can present asymptomatically, and develop persistent lymphocytosis or lymphosarcoma. The objective of this study was to estimate the prevalence and risk factors associated with bovine leukosis in Villavicencio, Colombia. Blood samples were taken from 636 animals, and obtained randomly from 24 herds. The samples were analysed using a Competition ELISA kit for the detection of anti-gp51 antibodies. Information on possible risk factors was collected, then OR and X2 were calculated, and statistically significant with p < 0.05 variables were included in a linear regression multivariate analysis. The general seroprevalence was 24.6% and the herd seroprevalence was 83.3%. The seroprevalence was 21.3% in males and 25.0% in females. The risk factors identified were abortion, non-bearing cows, artificial insemination, and use of common needles, Creole breed and participation in cattle exhibitions. The study confirmed the presence of bovine leukosis associated with reproductive and management factors.(AU)

A leucose bovina é causada por um vírus oncogênico do gênero Deltaretrovirus, causando prejuízos associados à queda dos indicadores produtivos e restrições à exportação de bovinos e derivados.Adoença tem um período de incubação prolongado entre 1 e 5 anos e os anticorpos podem ser detectados 2 a 3 semanas após a infecção. A doença pode se apresentar de forma assintomática, e evoluir para linfocitose persistente ou linfossarcoma. O objetivo do estudo foi estimar a prevalência e os fatores de risco associados à leucose bovina em Villavicencio, Colômbia. Amostras de sangue foram coletadas de 636 animais, obtidos aleatoriamente de 24 rebanhos.As amostras foram analisadas com o kit Competition ELISA para detecção de anticorpos anti-gp51. Foram coletadas informações sobre possíveis fatores de risco, se realizo um analise univariado entre as variáveis e a presença da seropositividad a leukosis bovina mediante o cálculo do OR e X2, as variáveis estatisticamente significativas com p<0,05 foram incluídas em uma análise multivariada de regressão linear. A soroprevalência geral foi de 24,6% e a soroprevalência do rebanho foi de 83,3%.Asoroprevalência foi de 21,3% em machos e 25,0% em fêmeas. Os fatores de risco identificados foram: aborto, vacas não reprodutivas, inseminação artificial e uso de agulha comum, raça crioula e exposições de gado. O estudo confirmou a presença de leucose bovina associada a fatores reprodutivos e de manejo.(AU)

Searching bovine papillomavirus presence in lesions seen on teats of cows

The samples were taken from 106 cows with various-looking lesions on their teats and ranged in age from 2 to 8 years. Enzyme-linked immunosorbent assay (ELISA) antigen (Ag) positive for the bovine papillomavirus (BPV) was found in 59 (55.7%) blood serum samples. PCR using FAP59/64 primers was positive for 24 (22.6%) samples. BPV-2 (40, 37.7%), BPV-6 (28, 26.4%), BPV-8 (30, 28.3%), BPV-9 (36, 34%), BPV-10 (32, 30.3%), and BPV-12 (22, 20.8%) were found in a PCR type-specific analysis of single and mixed type teat warts. The highest positivity was observed in BPV-2, BPV-9 and BPV-10 in flat and round forms, BPV-6, BPV-10, BPV-12, and mixed types in rice grain-cauliflower forms, BPV-9 and mixed types in filiform in the distribution of types based on the macroscopic appearance of teat lesions. As for the distribution of BPV types according to age, the most BPV-2 types were found in the age group of two years, the most BPV-10 types in the age group of three years, the most BPV-9 types in the age group of four years, the most BPV-8+BPV-12 types in the age group of five years, and the most mixed types between the ages of six and eight years. The existence of the virus was then checked using electron microscopy on the chosen samples (at least one investigation was conducted), and it was positively identified using BPV type-specific primers. The authors concluded that BPV detection using an ELISA (Ag) test from blood serum samples was shown to be less sensitive than BPV type-specific PCR from wart samples.

Bovine viral diarrhea virus infection in cattle - antioxidant status and some biochemical parameters

Background: Bovine viral diarrhea virus (BVDV) infections in cattle result in significant economic losses due to reproductive performance deficiencies caused by gastrointestinal, respiratory system infections, and transplacental infections. BVDV is one of the most important and widespread pathogens in cattle worldwide, including Turkey. Methods such as virus neutralization, enzyme-linked immunosorbent assay (ELISA), reverse transcriptase and polymerase chain reaction (RT-PCR) are used for the detection of the disease. The diagnosis of the disease in its subclinical form is challenging due to the lengthy and costly procedures involved. Investigating oxidative stress parameters in ruminants with various diseases contributes significantly to diagnosis and prognosis. This study aimed to investigate some oxidative stress and biochemical parameters in cattle infected with BVDV. Materials, Methods & Results: In the study, blood samples were collected from 80 Simmental breed cows aged between approximately 4 and 8 years to determine the presence of BVDV antibodies using the ELISA method. Based on the results obtained, study groups were organized. The study included a group of 10 animals with positive antibody levels as the infected group, and a group of 10 animals with negative antibody levels as the healthy group. Blood samples were taken from the animals, and serum separation was ensured. In the obtained serum samples, levels of vitamin E, vitamin A, ß-Carotene, catalase, GSH-Px, and MDA were determined using spectrophotometric methods. In addition, serum total protein, albumin, alkaline phosphatase (ALP), aspartate aminotransferase (AST), glucose, low-density lipoprotein (LDL), high-density lipoprotein (HDL), calcium (Ca), and phosphorus (P) were measured using commercial test kits and an autoanalyzer. In the study, it was observed that the differences in serum MDA, vitamin E, vitamin A, ß-carotene, and catalase levels were statistically significant between the healthy and BVDV-infected groups (P < 0.001). The activity of GSH-Px was also found to be statistically different between the groups (P < 0.01). Among the biochemical parameters, HDL, LDL, and AST levels were found to be statistically significant between the healthy and BVDV-infected groups (P < 0.001). Additionally, ALP and glucose levels were found to be statistically significant (P < 0.01). However, although there were differences in the levels of total protein, albumin, Ca, and P between the groups, these results were not statistically significant. Discussion: Although the diagnosis of the disease was partially made based on clinical observations in BVDV infections, the ELISA method was used for accurate diagnosis. Furthermore, it was found that there was a significant difference in MDA concentration between the healthy and infected groups, indicating oxidative damage caused by the virus. Similarly, significant differences in vitamin E, vitamin A, ß-carotene, GSH-Px, and catalase levels were observed between the groups, indicating a decrease in antioxidant values due to the infection. In addition, differences in ALP, AST, glucose, LDL, and HDL levels were found between the groups. This difference is thought to be related to the effects of the disease agent on the liver and systemically. This study demonstrates that, in addition to the viral pathogen, antioxidant and biochemical values are important criteria in the detection of the disease.

Oxidative activity of corpus luteum and ovarian parenchyma in Bos taurus indicus heifers

Abstract The aim of this study was to evaluate the oxidative stress in ovaries and corpus luteum (CL) of Bos taurus indicus females and the oxidant effect of CL in ovarian tissues in regions near, intermediate, or distant from it. Ovaries (n=12) of Nelore heifers (n=6) were collected from a slaughterhouse and fragmented. Experiment 1, each ovary was obtained from three fragments, resulting in 18 fragments of ovaries with CL (OV+CL) and another 18 fragments of ovaries without CL (OV-CL). Three fragments were generated from CL, totaling 18 CL fragments. In experiment 2, the ovarian fragments were removed from specific regions near, intermediate, or distant from the CL. All the fragments were placed in Eppendorf-type microtubes (1 mL), kept in a thermal container at 4 ºC, and then stored in a -80 ºC freezer for analysis of oxidative stress (TBARS and NBT) and antioxidant potential (FRAP and ABTS). In the antioxidant activity analysis, luteal tissues showed more antioxidant activity than ovarian tissue (FRAP = P < 0.0001; ABTS = P < 0.02). In the oxidative stress analysis, CL had lower levels of reactive oxygen species (ROS; TBARS = P < 0.03; NBT = P < 0.0001) than ovarian tissues. There was no difference in antioxidant activity and oxidative stress between the fragments obtained from different regions (OV+CL versus OV-CL; P > 0.05). The presence of CL in the ovaries of Bos taurus indicus females did not influence the oxidative stress or antioxidant potential of the gonad. Thus, the removal of ovarian fragments with or without the presence of CL indicates that biotechnologies such as in vitro follicle cultivation is possible.

Oxidative activity of corpus luteum and ovarian parenchyma in Bos taurus indicus heifers

The aim of this study was to evaluate the oxidative stress in ovaries and corpus luteum (CL) of Bos taurus indicus females and the oxidant effect of CL in ovarian tissues in regions near, intermediate, or distant from it. Ovaries (n=12) of Nelore heifers (n=6) were collected from a slaughterhouse and fragmented. Experiment 1, each ovary was obtained from three fragments, resulting in 18 fragments of ovaries with CL (OV+CL) and another 18 fragments of ovaries without CL (OV-CL). Three fragments were generated from CL, totaling 18 CL fragments. In experiment 2, the ovarian fragments were removed from specific regions near, intermediate, or distant from the CL. All the fragments were placed in Eppendorf-type microtubes (1 mL), kept in a thermal container at 4 ºC, and then stored in a -80 ºC freezer for analysis of oxidative stress (TBARS and NBT) and antioxidant potential (FRAP and ABTS). In the antioxidant activity analysis, luteal tissues showed more antioxidant activity than ovarian tissue (FRAP = P < 0.0001; ABTS = P < 0.02). In the oxidative stress analysis, CL had lower levels of reactive oxygen species (ROS; TBARS = P < 0.03; NBT = P < 0.0001) than ovarian tissues. There was no difference in antioxidant activity and oxidative stress between the fragments obtained from different regions (OV+CL versus OV-CL; P > 0.05). The presence of CL in the ovaries of Bos taurus indicus females did not influence the oxidative stress or antioxidant potential of the gonad. Thus, the removal of ovarian fragments with or without the presence of CL indicates that biotechnologies such as in vitro follicle cultivation is possible.(AU)

Espermiogênese alterada em um touro doador de sêmen: relato de caso / Altered spermiogenesis in a semen donor bull: case report

A espermiogênese é o processo final da espermatogênese no qual a espermátide se transforma em espermatozoide. Durante esse processo podem ocorrer alterações espermáticas, especialmente no acrossoma, na morfologia da cabeça, na condensação da cromatina e na formação dos vacúolos nucleares. A diferenciação entre os quadros clínicos é feita com repetições dos exames andrológicos. A avaliação morfológica indica a situação da espermiogênese nas 4 semanas anteriores a coleta do sêmen. Com os resultados do exame andrológico é possível identificar a qualidade seminal naquele período. O presente artigo é o relato de caso de um touro jovem doador de sêmen com 26 meses de idade, no início do regime de coletas de sêmen em um Centro de Coleta e Processamento de Sêmen no Sul do Brasil. A produção espermática deste animal foi avaliada durante 12 meses de coleta, apresentando sempre morfologia espermática muito alterada e motilidade média de 54% no exame imediato. Em 13 coletas seminais, o ejaculado deste animal apresentou em média 86% de defeitos maiores, 10% de defeitos menores e 96% de defeitos totais. Os defeitos maiores, que são os que possuem maior efeito na fertilidade, estão diretamente ligados a espermiogênese e os defeitos menores, que possuem menor efeito na fertilidade, são ocasionados principalmente durante o trânsito pelo epidídimo. O quadro clínico desse animal demonstrou a importância do exame morfológico para avaliar a espermatogênese e principalmente o processo de diferenciação final da espermátide em espermatozoide. Casos como este devem ser descritos como espermiogênese alterada e o reprodutor deve ser afastado da reprodução, após descartar alterações morfológicas devido a degeneração testicular grave.(AU)

Spermiogenesis is the final process of spermatogenesis in which the spermatid transforms into sperm. During this process, sperm alterations may occur, especially in the acrosome, head morphology, chromatin condensation and formation of nuclear vacuoles. Differentiation between clinical conditions is made with repetitions of andrological exams. The morphological evaluation indicates the status of spermiogenesis in the 4 weeks prior to semen collection. With the results of the breeding soundness examination, it is possible to identify the seminal quality in that period. The present article is a case report of a young 26-month-old semen donor bull, that started semen collection routines at a Semen Collection and Processing Center in southern Brazil. The sperm production of this animal was evaluated during 12 months of collection, always showing very altered sperm morphology and average motility of 54% in the immediate examination. In 13 seminal collections, the ejaculate of this animal presented an average of 86% of major defects, 10% of minor defects and 96% of total defects. Major defects, which have the greatest impact on fertility, are directly linked to spermiogenesis and minor defects, which have less effect on fertility, are mainly caused during transit through the epididymis. The clinical conditions of this animal demonstrated the importance of the morphological examination to evaluate spermatogenesis and especially the process of final differentiation of the spermatid into spermatozoa. Cases like this should be described as altered spermiogenesis and the bull should be withdrawn from breeding, after ruling out morphological changes due to severe testicular degeneration.(AU)

First detection of Trypanosoma vivax in dairy cattle from the northwest region of Minas Gerais, Brazil / Primeira detecção de Trypanosoma vivax em bovinos leiteiros da região noroeste de Minas Gerais, Brasil

A tripanossomíase bovina é causada pelo protozoário Trypanosoma vivax. A transmissão biológica ocorre apenas no continente africano pela mosca Tsé-tsé, de forma mecânica por dípteros hematófagos em todos os continentes, ou pelo compartilhamento de agulhas e por práticas associadas. O estudo teve como objetivo relatar o primeiro diagnóstico parasitológico, sorológico e molecular de T. vivax em bovinos leiteiros provenientes de cinco propriedades do município de Unai, Minas Gerais, Brasil. Cento e quinze animais selecionados por conveniência apresentavam sinais clínicos ou pertenciam a lotes de animais suspeitos. Foram detectados positivos pelos testes parasitológico (técnica de Woo), sorológico (ELISA) e molecular (LAMP). A maior prevalência global para T. vivax foi de 11,11% na propriedade A. O único sinal clínico dos animais positivos estudados foi baixa taxa de concepção. O primeiro diagnóstico de tripanossomíase no noroeste mineiro é extremamente importante, haja vista o tamanho do rebanho leiteiro da região e as possíveis perdas econômicas provocadas pela enfermidade. Ademais, faz-se necessário maior controle sanitário na região, uma vez que a transmissão no Brasil é intimamente ligada às práticas de compartilhamento de agulhas no manejo dos animais e ao parasitismo de moscas hematófagas.

Natural association between bovine leukemia virus and reproductive infectious diseases

Background: Enzootic bovine leukosis (EBL) is a widespread infectious disease caused by the bovine leukemia virus (BLV), which results in immune system dysfunction. The resulting immunosuppression may lead to an increased prevalence of other diseases. Dairy cows infected have altered immune function associated with decreased milk production and shortened lifespan and decreased immune response to immunization. BLV infection, however, is often asymptomatic, so any connection between subclinical infection and common reproductive diseases remains unknown. This study aimed to describe the relationship between naturally occurring subclinical BLV and infectious reproductive diseases seroconversion in the field. Materials, Methods & Results: The diseases investigated included Bovine viral diarrhoea (BVD), Bovine alfaherpesvirus 1 (BoHV-1), Bovine gammaherpesvirus 4 (BoHV-4), Chlamydiosis, Leptospirosis, Brucellosis and Neosporosis in dairy cattle. Six hundred fifty-five sera samples from the northern and south-central regions of Uruguay, from asymptomatic female Holstein and Holstein crosses without a history of vaccination against reproductive diseases were processed using reference diagnostic methods (Seronautalization, ELISA, MAT, Rose Bengal Plate test). The seroprevalence of BLV was 20.0%. Seroprevalence of reproductive diseases BVD, BoHV-1 and BoHV-4 were 99.3%, 41.2% and 27.3% of the populations, respectively, and the total seroprevalence of Leptospirosis, bovine Neospora caninum and Chlamydiosis were 19.8%, 29.8% and 33.0% respectively. The results revealed positive associations between naturally contracted BLV and the presence of antibodies against BoHV-1 (P = 0.002), as well as between naturally contracted BLV and presence of antibodies against Leptospira spp. (P = 0.028). Discussion: BLV infection can impact innate and adaptive immune system cells and alter the proper functioning of uninfected cells. BLV infection may also induce changes in the complex balance of cytokine expression, cell proliferation, and programmed cell death in T- and B-lymphocytes, which is critical for immune competence and effective response to infectious challenges. The progression of BLV infection has a substantial effect on host defense mechanisms. Indeed, lowmagnitude serologic responses to a commercial foot-and-mouth disease vaccine and a J5 Escherichia coli vaccine have been observed. These results are supported by recent trial studies showing a reduced immune response to vaccination against BoHV-1 and Leptospira spp. in asymptomatic animals infected with BLV. These are 2 of the most prevalent infectious reproductive diseases in cattle worldwide, and our results provide evidence that a link between BLV and susceptibility to these diseases may exist. Although there is evidence of the co-occurrence of these diseases, it remains unknown whether there is a direct or indirect effect of BLV on infertility, embryonic loss, or abortion. Another possibility is that natural infection with these reproductive pathogens (BoHV-1, Leptospira, or others) promotes BLV expression, negatively affecting the farms where these pathogens are endemic. Considering the high seroprevalence of BLV in dairy herds in North and South America where the infection is endemic, it was explored BLV's role as an immunosuppressant by quantifying its co-occurrence with diseases that affect reproductive performance in breeding herds. Future work should clarify the role of BLV and the co-occurring pathogens in causing infertility or abortions.

Embryo biopsies for genomic selection in tropical dairy cattle

Genomic selection has transformed the livestock industry, enabling early-life selection of animals. Biopsy sampling of pre-implantation embryos has been described since 1968. However, it was only after 2010, with the advancement of molecular biology techniques such as whole genomic amplification and SNP Chips, that next-generation sequencing became commercially available for bovine embryos. It is now possible to make decisions about which embryos to transfer not only based on recipients' availability or embryo morphology but also on genomic estimates. This technology can be implemented for a wide spectrum of applications in livestock. In this review, we discuss the use of embryo biopsy for genomic selection and share our experience with Gir and Girolando Brazilian breeding programs, as well as future goals for implementing it in Brazilian bovine in vitro embryo production practices.(AU)

Emulsifier in Diets with Different Alternative Lipid Sources: Effects on the Health of Japanese Quails

The present research aims to evaluate the inclusion of an emulsifying additive based on mono- and diglycerides of fatty acids in diets with different lipid sources, studying its effects on duodenum histology, biometry of the digestive and reproductive tract, and blood biochemical profile of Japanese quails. The study was carried out in the experimental aviary of the Federal University of Grande Dourados, Mato Grosso do Sul, Brazil. The experimental design was completely randomized, with diets in a 3x2 factorial design with three different lipid sources (soybean oil, poultry fat, and beef tallow), either supplemented or not with an emulsifier. The birds that received a diet composed of poultry fat and an emulsifying additive had a larger gizzard. Animals that received a diet containing an emulsifier, regardless of the lipid source, had greater heart weight. The duodenum of animals that received bovine tallow in their diet showed a greater height and villus:crypt ratio. Biochemical parameters were not affected by either the lipid source or the inclusion of emulsifier in quail diets. The biochemical blood profile of birds in treatments did not show significant changes among all diets used. The use of soybean oil, bovine tallow, and poultry fat with or without the inclusion of an emulsifier does not show changes in the health and development of the quails' organs, which also indicates that the health of Japanese quails in the laying phase is kept in good standards.(AU)

Mechanisms of acaricide resistance in ticks

Background: In several countries, including Brazil, the livestock industry plays a key role in the country's economy. Brazil has the second largest bovine herd in the world and the biggest commercial herd. Ticks are an ongoing problem for both large operation cattle producers and small family farmers. Rhipicephalus microplus causes expressive losses in cattle breeding, since it occurs in important beef production zones like South America, Africa, and Oceania. Some of the negative consequences of tick infestation to cattle breeding are anemia, loss in milk and beef production, and transmission of Babesia bovis and B. bigemina. Significant losses are caused by the cattle tick (R. microplus) in several regions of the world, costing around US$ 3.3 billion per year to the Brazilian livestock industry alone. The tick control methods are mainly based on synthetic acaricides. However, the improvement of current tick control requires the identification of new molecular targets in tick physiology and development of molecule compounds to target important physiology pathways. The strategies proposed to address this issue are expand the knowledge about the molecules involved in the detoxification of chemicals to enhance the efficacy of the acaricides as well as to develop new compounds for chemical control. Review: Tick control is currently based on chemical acaricides; however, effective control and prevention of tick infestation remain distant goals. In recent decades, a progressive decrease in the efficiency of acaricides due to drug resistance has been observed. Acaricide resistance is an evolutionary adaptation, which implies the existence of behavioral and physiological mechanisms that allow the survival of resistant individuals. Four resistance mechanisms are described: behavioral resistance, reduced drug penetration, target site insensitivity and increased drug detoxification. Augmented drug detoxification may be due to increased activity of enzymes or transporters due to increased gene expression or mutations in some genes. Research focus on mechanisms of acaricide resistance in ticks characterized detoxification pathways based on (1) increased activity of enzymes (cytochrome p450, esterase and GST) which play a role in biochemically altering acaricides towards decreased toxicity and, (2) enhanced excretion of the modified less toxic compounds. To bypass the current problems, a better understanding of the biology, physiology, and molecular biology of the mechanisms of resistance to acaricides is fundamental to prolong their efficiency in controlling ticks. Moreover, identifying the genes and proteins associated with resistance can support in the development of more sensitive diagnostic methods to identify acaricide resistance, as well as improving control strategies. Discussion: In the last years, many researchers have been studying resistance mechanisms and important advances have been made which showed that, in several tick species, ABC transporters, esterases, P-450 cytochromes and glutathione-S-transferases participate in acaricide resistance. The characterization of the alterations in the targets in tick physiology and identification of new drugs with potential to tick control are crucial goals to increase tick control

Abordagem terapêutica hormonal para cistos foliculares ovarianos em vacas leiteiras - relato de casos / Hormonal therapeutic approach for ovarian follicular cysts in dairy cattle - case reports / Enfoque terapéutico hormonal para quistes foliculares ováricos en vacas lecheras - informe de casos

Vacas de alta produção de leite estão mais sujeitas a desequilíbrios metabólicos pós-parto, influenciando na performance reprodutiva subsequente. Alterações relacionadas a funcionalidade do ovário podem desencadear desequilíbrios hormonais e desordens reprodutivas, como a formação de cistos ovarianos, os quais podem interferir no processo de ovulação e impactar substancialmente na eficiência reprodutiva. Neste estudo de caso em específico, sete vacas leiteiras foram diagnosticadas com a presença de cistos foliculares ovarianos. Foram submetidas a terapia hormonal envolvendo o uso de análogos de Hormônio Liberador de Gonadotrofinas (GnRH) e prostaglandina-F2a (PGF-2a), a qual apresentou resultados satisfatórios. A abordagem terapêutica hormonal tem sido amplamente utilizada devido à sua efetividade na resolução dos cistos ováricos e melhoria da função reprodutiva. É fundamental que o médico veterinário tenha conhecimento clínico sobre a abordagem adequada de cistos ovarianos, a fim de proporcionar saúde e bem-estar aos animais, garantindo bons índices em produção de leite e desempenho reprodutivo.

High milk production cows are more likely to postpartum metabolic imbalances, which may influence subsequent reproductive performance. Changes in ovarian functionality can trigger hormonal alterations and reproductive disorders, such as the formation of ovarian cysts, which can interfere with the ovulation process and substantially impact reproductive efficiency. In this specific case study, seven dairy cows were diagnosed with the presence of ovarian follicular cysts. They were submitted to hormonal therapy involving the use of Gonadotropin-Releasing Hormone (GnRH) and prostaglandin F2a (PGF-2a) analogs, which presented satisfactory results. Hormonal therapeutic approaches have been widely employed due to their effectiveness in resolving ovarian cysts and improving reproductive function. It is essential for the veterinarian to have clinical knowledge about the appropriate management of ovarian cysts in order to provide the health and welfare of the animals, ensuring suitable rates in milk production and reproductive performance.

Las vacas de alta producción de leche tienen mayor probabilidad de desequilibrios metabólicos posparto, lo cual puede influir en el rendimiento reproductivo posterior. Los cambios en la funcionalidad ovárica pueden desencadenar alteraciones hormonales y trastornos reproductivos, como la formación de quistes ováricos, que pueden interferir en el proceso de ovulación y afectar sustancialmente la eficiencia reproductiva. En este estudio de caso específico, se diagnosticó la presencia de quistes foliculares ováricos en siete vacas lecheras. Se sometieron a terapia hormonal que incluyó el uso de análogos de Hormona Liberadora de Gonadotropinas (GnRH) y prostaglandina F2α (PGF-2a), lo cual arrojó resultados satisfactorios. Los enfoques terapéuticos hormonales se han utilizado ampliamente debido a su efectividad en la resolución de quistes ováricos y la mejora de la función reproductiva. Es fundamental que el médico veterinario tenga conocimiento clínico sobre el manejo adecuado de los quistes ováricos para garantizar la salud y el bienestar de los animales, asegurando adecuadas tasas de producción de leche y rendimiento reproductivo.

Effects of autolyzed yeast on leukocyte oxidative metabolism and pneumonia occurrence in feedlot steers / Efeito da levedura autolisada no metabolismo oxidativo leucocitário e na ocorrência de pneumonia em novilhos terminados em confinamento

Although yeast supplementation has shown an immunostimulant effect in feedlot cattle, the question remains whether it influences the occurrence of the bovine respiratory disease complex (BRD) in feedlot cattle. Therefore, our objective was to evaluate whether dietary supplementation with autolyzed yeast affects the efficiency of blood phagocytes, reducing inflammation and the occurrence of BRD in feedlot cattle. A randomized experimental trial was conducted with whole steers with half Angus blood for 106 days: control (n=10): diet without yeast; Y4 (n=12): yeast diet (4g per animal per day); and Y7 (n=7): yeast diet (7g per animal per day). On Days 0, 14, 42, 70, and 105, blood count, leukocyte oxidative metabolism, serum haptoglobin, frequency of nasal secretion and orbital temperature were evaluated. On slaughter day, a lung evaluation was performed. On Days 42 and 70 of feedlot finishing, the treated animals showed increased leukocyte oxidative metabolism (D42 P=0.04; D70 P=0.02) compared to the control group. These animals had a lower frequency of mucopurulent nasal secretion, lower orbital temperature and reduced pneumonia occurrence than the treated groups (P=0.05) with less lung lesion severity (P=0.01), allowing us to infer that the autolyzed yeast improves blood phagocytic function and decreases the inflammation and BRD of feedlot steers, especially at a dose of 4g.

Embora a suplementação com leveduras tenha demonstrado efeito imunoestimulante em bovinos, permanece-se a dúvida se ela influencia na ocorrência do complexo doença respiratória bovina (CDRB) em bovinos confinados. Portanto, objetivou-se avaliar se a suplementação dietética com levedura autolisada influencia a eficiência de fagócitos sanguíneos, reduzindo inflamações e a ocorrência de CDRB em bovinos confinados. Realizou-se um ensaio experimental randomizado com 36 novilhos inteiros 1/2 sangue Angus por 106 dias: controle (n=10): dieta sem levedura; Y4 (n=12): dieta com levedura (4g por animal por dia); e Y7 (n=7): dieta com levedura (7g por animal por dia). Nos dias 0, 14, 42, 70 e 105, hemograma, metabolismo oxidativo de leucócitos, haptoglobina sérica, frequência de secreção nasal e temperatura orbital foram avaliados. No dia do abate, foi realizada avaliação pulmonar. Nos dias 42, e 70 de terminação em confinamento, os animais tratados apresentaram aumento do metabolismo oxidativo dos leucócitos (D42 P=0,04; D70 P=0,02) em relação ao grupo controle. Esses animais apresentaram menor frequência de secreção nasal mucopurulenta, menor temperatura orbital, menor ocorrência de pneumonias (P=0,05) com menor gravidade das lesões pulmonares (P=0,01), permitindo inferir que a levedura autolisada na dieta melhorou a eficiência dos fagócitos sanguíneos, reduzindo inflamações e a ocorrência de CDRB, especialmente na dosagem de 4g.

A glycoprotein E gene-deleted bovine alphaherpesvirus 1 strain is attenuated and immunogenic for calves with passive immunity upon intranasal immunization / Cepa de alfaherpesvírus bovino 1 com deleção do gene da glicoproteína E é atenuada e imunogênica para bezerros com imunidade passiva após imunização intranasal

ABSTRACT: Vaccination has been used to prevent the losses associated with Bovine alphaherpesvirus 1 (BoHV-1) infection but passively acquired antibodies may compromise vaccine efficacy. Intranasal immunization (IN) of calves with modified live viral BoHV-1 vaccines has proven to overcome the acquired passive antibodies and confer adequate protection. Herein, we evaluated the safety and immunogenicity of a glycoprotein E-deleted Brazilian BoHV-1 strain (BoHV-1gEΔ) for IN immunization of calves. Ten 1-to-2 months-old calves with virus-neutralizing titers (VN) ranging from 2-64 were immunized IN with viable BoHV-1gEΔ (107.1 TCID50) and four remained as unvaccinated controls (VN titers 8-32). After IN immunization, calves presented a transient (2-6 days) mild nasal secretion and shed the vaccine virus in nasal secretions in low titers (<102.6TCID50/mL) for 4-8 days. Interestingly, the vaccinated calves did not show an increase in VN titers after vaccination. Rather, they presented a gradual reduction in serum VN antibodies in the following weeks - similarly to unvaccinated controls. Upon IN challenge with a virulent heterologous BoHV-1 strain at day 55 post-immunization (107.63TCID50), vaccinated calves shed significantly less virus from day 6 post-challenge onwards (p < 0.07) and for a shorter period of time than the controls (p < 0.0024). Importantly, both the duration and intensity of clinical signs were reduced in vaccinated animals. In addition, vaccinated calves showed an abrupt raise in VN titers post-challenge, indicating adequate immunological priming by vaccination. In summary, immunization of calves harboring passive antibodies with BoHV-1gEΔ by the IN route was able to prime the immunity to afford partial virological and clinical protection upon challenge.

RESUMO: A vacinação tem sido usada para prevenir perdas associadas à infecção pelo alfaherpesvírus bovino 1 (BoHV-1), embora anticorpos adquiridos passivamente possam comprometer a eficácia das vacinas. A imunização intranasal (IN) de bezerros com vacinas de BoHV-1 vivas modificadas pode contornar o obstáculo relacionado à presença de anticorpos adquiridos passivamente, conferindo proteção aos animais vacinados. Nesse contexto, avaliou-se a segurança e imunogenicidade de uma cepa brasileira de BoHV-1 com deleção no gene da glicoproteína E (BoHV-1gEΔ) na imunização IN de bezerros. Dez bezerros, de um a dois meses de idade e com títulos neutralizantes (VN) variando de 2-64, foram inoculados IN com BoHV-1gEΔ (107,1TCID50), e quatro permaneceram como controles não vacinados (títulos de VN 8-32). Após a instilação IN, os bezerros apresentaram secreção nasal transitória leve (2-6 dias) e excretaram o vírus vacinal nas secreções nasais em baixos títulos (<102,6TCID50/mL) por 4-8 dias. Interessantemente, os bezerros vacinados não apresentaram aumento nos títulos de anticorpos neutralizantes após a vacinação. Em vez disso, eles apresentaram uma redução gradual nos anticorpos neutralizantes séricos nas semanas seguintes - semelhante aos controles não vacinados. Após o desafio IN com uma cepa BoHV-1 virulenta heteróloga no dia 55 pós-imunização (107,63TCID50), os bezerros vacinados excretaram o vírus em títulos menores a partir do sexto dia pós-desafio (p < 0,07) e por um período de tempo menor do que o observado nos controles (p < 0,0024). É importante notar que tanto a duração quanto a intensidade dos sinais clínicos foram reduzidas nos animais vacinados. Além disso, os bezerros vacinados apresentaram um aumento abrupto nos títulos neutralizantes após o desafio, indicando uma imunização adequada por BoHV-1gEΔ. Em resumo, a imunização IN de bezerros com anticorpos passivos com a cepa BoHV-1gEΔ foi capaz de estimular a imunidade, proporcionando proteção virológica e clínica parciais após o desafio.

A glycoprotein E gene-deleted bovine alphaherpesvirus 1 strain is attenuated and immunogenic for calves with passive immunity upon intranasal immunization / Cepa de alfaherpesvírus bovino 1 com deleção do gene da glicoproteína E é atenuada e imunogênica para bezerros com imunidade passiva após imunização intranasal

Vaccination has been used to prevent the losses associated with Bovine alphaherpesvirus 1 (BoHV-1) infection but passively acquired antibodies may compromise vaccine efficacy. Intranasal immunization (IN) of calves with modified live viral BoHV-1 vaccines has proven to overcome the acquired passive antibodies and confer adequate protection. Herein, we evaluated the safety and immunogenicity of a glycoprotein E-deleted Brazilian BoHV-1 strain (BoHV-1gEΔ) for IN immunization of calves. Ten 1-to-2 months-old calves with virus-neutralizing titers (VN) ranging from 2-64 were immunized IN with viable BoHV-1gEΔ (107.1 TCID50) and four remained as unvaccinated controls (VN titers 8-32). After IN immunization, calves presented a transient (2-6 days) mild nasal secretion and shed the vaccine virus in nasal secretions in low titers (<102.6TCID50/mL) for 4-8 days. Interestingly, the vaccinated calves did not show an increase in VN titers after vaccination. Rather, they presented a gradual reduction in serum VN antibodies in the following weeks - similarly to unvaccinated controls. Upon IN challenge with a virulent heterologous BoHV-1 strain at day 55 post-immunization (107.63TCID50), vaccinated calves shed significantly less virus from day 6 post-challenge onwards (p < 0.07) and for a shorter period of time than the controls (p < 0.0024). Importantly, both the duration and intensity of clinical signs were reduced in vaccinated animals. In addition, vaccinated calves showed an abrupt raise in VN titers post-challenge, indicating adequate immunological priming by vaccination. In summary, immunization of calves harboring passive antibodies with BoHV-1gEΔ by the IN route was able to prime the immunity to afford partial virological and clinical protection upon challenge.

A vacinação tem sido usada para prevenir perdas associadas à infecção pelo alfaherpesvírus bovino 1 (BoHV-1), embora anticorpos adquiridos passivamente possam comprometer a eficácia das vacinas. A imunização intranasal (IN) de bezerros com vacinas de BoHV-1 vivas modificadas pode contornar o obstáculo relacionado à presença de anticorpos adquiridos passivamente, conferindo proteção aos animais vacinados. Nesse contexto, avaliou-se a segurança e imunogenicidade de uma cepa brasileira de BoHV-1 com deleção no gene da glicoproteína E (BoHV-1gEΔ) na imunização IN de bezerros. Dez bezerros, de um a dois meses de idade e com títulos neutralizantes (VN) variando de 2-64, foram inoculados IN com BoHV-1gEΔ (107,1TCID50), e quatro permaneceram como controles não vacinados (títulos de VN 8-32). Após a instilação IN, os bezerros apresentaram secreção nasal transitória leve (2-6 dias) e excretaram o vírus vacinal nas secreções nasais em baixos títulos (<102,6TCID50/mL) por 4-8 dias. Interessantemente, os bezerros vacinados não apresentaram aumento nos títulos de anticorpos neutralizantes após a vacinação. Em vez disso, eles apresentaram uma redução gradual nos anticorpos neutralizantes séricos nas semanas seguintes - semelhante aos controles não vacinados. Após o desafio IN com uma cepa BoHV-1 virulenta heteróloga no dia 55 pós-imunização (107,63TCID50), os bezerros vacinados excretaram o vírus em títulos menores a partir do sexto dia pós-desafio (p < 0,07) e por um período de tempo menor do que o observado nos controles (p < 0,0024). É importante notar que tanto a duração quanto a intensidade dos sinais clínicos foram reduzidas nos animais vacinados. Além disso, os bezerros vacinados apresentaram um aumento abrupto nos títulos neutralizantes após o desafio, indicando uma imunização adequada por BoHV-1gEΔ. Em resumo, a imunização IN de bezerros com anticorpos passivos com a cepa BoHV-1gEΔ foi capaz de estimular a imunidade, proporcionando proteção virológica e clínica parciais após o desafio.

Efficacy assessment of an intramammary formulation based on soluble polypyrrole in cows with experimentally induced mastitis / Avaliação da eficácia de uma formulação intramamária à base de polipirrol solúvel em vacas com mastite experimentalmente induzida

Considerando a necessidade de terapias alternativas para a mastite bovina, uma inflamação, normalmente de causa infecciosa, de alta prevalência e de alto impacto econômico nas fazendas leiteiras, avaliou-se a eficácia antimicrobiana, in vivo, de uma formulação intramamária à base de polipirrol (PPy) solúvel, um polímero condutor promissor para aplicações biomédicas, especialmente como potente antibacteriano. Neste ensaio, quartos mamários de vacas holandesas sadias (n = 8) foram inoculados com Staphylococcus aureus e tratados, por via intramamária, com formulação experimental à base de PPy solúvel (5%) e com formulação comercial à base de sulfato de gentamicina. O efeito desses tratamentos foi avaliado com a realização de lactoculturas, da Contagem Bacteriana Total (TBC), da Contagem de Células Somáticas (SCC) e da análise da composição do leite das amostras obtidas de quartos mamários, em sete momentos experimentais. Avaliação hematológica dos animais também foi realizada. A aplicação intramamária de três doses da formulação experimental à base de PPy solúvel resultou em maiores log/mL da TBC e da SCC quando comparadas ao grupo controle positivo e ao grupo que recebeu sulfato de gentamicina. A administração da formulação experimental não induziu alterações na composição do leite e nos parâmetros hematológicos. Alguns fatores farmacocinéticos e farmacodinâmicos do PPy solúvel podem ser atribuídos a ineficácia antimicrobiana da pomada experimental. Outras pesquisas devem ser realizados em prol do desenvolvimento de formulações que permitam a atuação antibacteriana do PPy solúvel no ambiente intramamário de vacas leiteiras.

Given the need for alternative therapies for bovine mastitis, an infectious disease of high prevalence and significant economic impact in dairy farms, this study evaluated the in vivo antimicrobial efficacy of an intramammary formulation based on soluble polypyrrole (PPy), a promising conductive polymer for biomedical applications, especially as an effective antimicrobial agent. In this assay, mammary quarters of healthy Holstein Friesian cows (n = 8) were inoculated with Staphylococcus aureus and treated, via the intramammary route, with an experimental formulation based on soluble PPy (5%) and a commercial formulation based on gentamicin sulfate. The effect of these treatments was evaluated based on milk cultures, Total Bacterial Count (TBC), Somatic Cell Count (SCC), and milk composition analysis of mammary quarter samples in seven experimental moments. In addition, hematological evaluations of the animals were also performed. The intramammary application of three levels of the experimental formulation based on soluble PPy resulted in higher log/mL of the TBC and SCC compared to a positive control group and a group that received gentamicin sulfate. The experimental formulation did not induce changes in milk composition or in hematological parameters. Certain factors related to pharmacokinetics, such as the type of carrier used in the formulation and the pharmacodynamics of soluble PPy, may have contributed to the antimicrobial ineffectiveness of the experimental formulation. The results of this study neither define nor decrease the antimicrobial potential of soluble PPy. Further research is required to develop formulations that enable the antibacterial action of soluble PPy in the intramammary environment of dairy cows.

Sarcocystis spp. detection in cattle using different diagnostic methods / Detecção de Sarcocystis spp. em bovinos usando diferentes métodos de diagnóstico

Cattle are considered intermediate hosts of Sarcocystis, which can cause clinical signs and lower performance in the acute phase of infection. Sarcocystis spp. are usually not visible to the naked eye during the post mortem inspection. Moreover, fresh microscopic examination and transmission electron microscopy techniques are difficult to apply to large samples. Therefore, extensive studies on Sarcocystis infection in cattle using molecular and serological methods are required. Here, we investigated Sarcocystis spp. infection in cattle using fresh microscopic examination and polymerase chain reaction of myocardium samples and compared the results with the presence of antibodies against Sarcocystis spp. in corresponding serum samples detected using indirect fluorescent antibody test. Microscopic Sarcocystis were observed in 100% of the myocardial samples, and Sarcocystis DNA was present in 86% (43/50) of these samples. Antibodies against Sarcocystis spp. were detected in 96% (48/50) and 80% (40/50) of the serum samples at 1:25 and 1:200 dilutions, respectively. The three associated methods (fresh microscopic examination, PCR and serology) showed good sensitivity and detection for Sarcocystis spp. compared with fresh microscopic examination (only), and they may facilitate diagnosis in live animals on a large scale as well as monitoring of the herd status.

Os bovinos são considerados hospedeiros intermediários de Sarcocystis, podendo causar sinais clínicos e menor desempenho na fase aguda da infecção. Sarcocystis spp. geralmente não são visíveis a olho nu durante a inspeção post mortem. Além disso, o exame microscópico a fresco e as técnicas de microscopia eletrônica de transmissão são difíceis de aplicar a uma amostras de grande tamanho. Portanto, são necessários extensos estudos sobre a infecção por Sarcocystis em bovinos usando métodos moleculares e sorológicos. Aqui, investigamos a infecção de Sarcocystis spp. em bovinos por meio de exame microscópico a fresco e reação em cadeia da polimerase de amostras de miocárdio e comparado os resultados com a presença de anticorpos contra Sarcocystis spp. em amostras de soro correspondentes detectadas usando o teste de anticorpos fluorescentes indiretos. Sarcocistos microscópicos foram observados em 100% das amostras de miocárdio, e o DNA de Sarcocystis estava presente em 86% (43/50) dessas amostras. Anticorpos contra Sarcocystis spp. foram detectados em 96% (48/50) e 80% (40/50) das amostras de soro nas diluições 1:25 e 1:200, respectivamente. Os três métodos associados (exame microscópico a fresco, PCR e sorologia) mostraram boa sensibilidade e detecção para Sarcocystis spp. em comparação com o exame microscópico fresco (apenas) e podem facilitar o diagnóstico em animais vivos em larga escala, bem como o monitoramento do status do rebanho.

Impact of persistent bovine viral diarrhea virus infection on indicators of innate and adaptive immune function in holstein calves and cows / Impacto da infecção persistente pelo vírus da diarreia viral bovina sobre indicadoresde função imunológica inata e adaptativa em bezerras e vacas holandesas

ABSTRACT: We evaluated some indicators of innate and humoral immune response in persistently infected (PI) Holstein calves and cows from 1 to 36 months of age matched with controls from the same herd. The effects were cataloged by grouping animals into the following age groups: <12 months, 13 to 24 months, and 25 to 36 months of age. Blood samples were collected once from each animal to measure total serum protein, haptoglobin, and neutralizing antibodies titers induced by respiratory virus vaccination. Total serum protein (g/dL) was lowest in PI calves younger until 24 months old, while haptoglobin concentration was higher in PI cattle. The serum neutralizing titers against BVDV and BRSV were lower in all PI calves and cattle than in controls. PI cattle have a high serum concentration of haptoglobin, and its possible dysregulated innate immune response appears to impact the efficacy of their adaptative immune responses, resulting in poor vaccine responsiveness.

RESUMO: O objetivo desta pesquisa foi avaliar alguns indicadores da resposta imune inata e humoral em bezerros a vacas persistentemente infectadas, entre um a 36 meses de idade, pareados com controles oriundos de um mesmo rebanho. As variáveis respostas foram avaliadas agrupando-se os animais nos seguintes grupos etários: < 12 meses, 13 a 24 meses, 25 a 36 meses de idade. Amostras sanguíneas foram coletadas para mensurar as concentrações séricas de proteína, haptoglobina e anticorpos neutralizantes induzidos pela vacinação contra as viroses respiratórias. Os teores de proteína sérica total (g/dL) foram menores nos animais persistentemente infectados (PI) jovens até 24 meses de idade, enquanto que a concentração de haptoglobina foi maior nos animais PI mais velhos (25 a 36 meses). Os títulos de anticorpos neutralizantes contra o BVDV e BRSV foi menor nos animais PIs independentemente da idade, comparado com o grupo controle. Os valores reduzidos ou nulos de anticorpos contra as viroses respiratórias, combinado com a evidência de resposta imune inata desregulada, contribui com a susceptibilidade dos animais PIs para as infecções secundárias.