Use of knowledge regarding LH receptors to improve superstimulatory treatments in cattle

Embryo transfer is a biotechnology that has been used worldwide to increase the production of offspring from female bovines. Treatments to induce multiple ovulations (superovulation) have evolved from superstimulatory protocols that depended upon detection of oestrus to treatments that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-timed AI (FTAI). The protocols associated with FTAI facilitate animal handling and produce at least as many viably embryos as conventional treatment protocols that required detection of oestrus. Recent knowledge regarding LH receptors (LHR) and follicular development can be applied to improve embryo transfer protocols. In fact, improvements in the superstimulatory treatment called the 'P-36 protocol', which include hormones that stimulate LHR, indicate that adjustments related to LHR availability may increase bovine embryo yield compared with conventional protocols based on the detection of oestrus.

Expression of protease nexin-1 and plasminogen activators during follicular growth and the periovulatory period in cattle

Extracellular matrix remodeling occurs during ovarian follicular development, mediated by plasminogen activators (PAs) and PA inhibitors including protease nexin-1 (PN-1). In the present study we measured expression/activity of the PA system in bovine follicles at different stages of development by timed collection of ovaries during the first follicular wave and during the periovulatory period, and in follicles collected from an abattoir. The abundance of mRNA encoding PN-1, tissue-type PA (tPA), urokinase (uPA) and PA inhibitor-1 (PAI-1) were initially upregulated by human chorionic gonadotropin (hCG) in bovine preovulatory follicular wall homogenates. PN-1, PAI-1 and tPA mRNA expression then decreased near the expected time of ovulation, whereas uPA mRNA levels remained high. PN-1 concentration in follicular fluid (FF) decreased and reached the lowest level at the time of ovulation, whereas plasmin activity in FF increased significantly after hCG. Follicles collected from the abattoir were classified as non-atretic, early-atretic or atretic based on FF estradiol and progesterone content: PN-1 protein levels in FF were significantly higher in non-atretic than in atretic follicles, and plasmin activity was correspondingly higher in the atretic follicles. No changes in PN-1 levels in FF were observed during the growth of pre-deviation follicles early in a follicular wave. These results indicate that PN-1 may be involved in the process of atresia in non-ovulatory dominant follicles and the prevention of precocious proteolysis in periovulatory follicles.

FGF10 inhibits dominant follicle growth and estradiol secretion in vivo in cattle

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Host specificity of sheep and cattle nematodes in São Paulo state, Brazil

The trial was carried out to investigate parasite host specificity and to analyse the dynamics of infection with nematodes parasitizing sheep and catt:le raised together or separately in São Paulo state, Brazil, and, also to clarify doubts about the systematics of species of the genus Haemonchus on the basis of cytological and morphological studies. Ten steers and 32 ewes were randomly assigned to three paddocks (P), as follows: P1, 5 steers; P2, 5 steers and 16 ewes; and P3, 16 ewes. The animals remained on these paddocks in continuous grazing throughout the trial (1-yr period). Faecal exams and larvae counting on pasture were performed fortnightly. Once a month two tracer lambs were placed in each paddock, while two tracer calves were also placed, but only in the eighth month of the trial. All these animals were slaughtered for worm identification and counting. At the end of the trial, one steer and one ewe from P2, which showed high faecal egg counts, were also slaughtered for the same purpose. Nematodes identified cytogenetically as H. placei presented spicule hooks longer than those identified as H. contortus. The following distribution of parasites in cattle and sheep was observed: Bunostomum phlebotomum, H. similis, Mammomonogamus laryngeus strongly adapted to cattle, H. placei and Cooperia punctata more adapted to cattle than to sheep, Trichostrongylus axel and C. spatulata apparently more adapted to cattle, T. colubriformis strongly adapted to sheep, H. contortus more adapted to sheep than to cattle and C. curticei apparently more adapted to sheep. Cross-infection was shown to occur involving some species, however, with time the animals apparently eliminate the species that are not well adapted to them. Therefore, grazing management systems using cattle and sheep appear to be promising for worm control in southeastern Brazil. (C) 1997 Elsevier B.V. B.V.

Field study on nematode resistance in Nelore-breed cattle

The present study evaluated Nelore cattle with different degrees of resistance to natural infections by gastrointestinal nematodes. One hundred weaned male cattle, 11-12 months of age, were kept on the same pasture and evaluated from October 2003 to February 2004. Faecal and blood samples were collected for parasitological, haematological and immunological tests. In February 2004, the 10 most resistant and the 10 most susceptible animals were selected based on individual means of nematode faecal egg counts (FEC). Such animals were slaughtered for worm burden determination and nematode species identification. The repeatability estimates for FEC (+/- S.D.), log-transformed FEC and packed-cell volume (PCV) in all animals were 0.3 (+/- 0.05), 0.26 (+/- 0.04) and 0.42 (+/- 0.05), respectively. The resistant group showed lower FEC and worm burdens than the susceptible group (P < 0.05). There were no significant differences between groups regarding mean body weight, weight gain, PCV and total serum protein values (P > 0.05). The resistant group showed higher total serum IgE levels (P < 0.05) and higher mean eosinophil blood counts. However, the latter was statistically significant only 42 days after the beginning of the study. Nematodes Cooperia punctata and Haemonchus placei were predominant and the correlation between Cooperia and Haemonchus burdens was 0.64 (P < 0.05), which indicated that animals presenting increased numbers of one of those genera probably had increased numbers of the other. The current study provides further evidence of IgE active role in nematode immunity and suggests that total serum IgE level might serve as an additional marker to select Nelore cattle that are responsive to H. placei and C. punctata infections. (C) 2007 Elsevier B.V. All rights reserved.

Anthelmintic resistance in cattle nematodes in northwestern São Paulo state, Brazil

The present study was carried out in the northwestern region of São Paulo State, Brazil, to determine the anthelmintic resistance status in cattle naturally infected with gastrointestinal nematodes. The anthelmintics tested were levamisole phosphate (Ripercol (R), Fort Dodge), albendazole sulphoxide (Ricobendazole, Fort Dodge), ivermectin (Ivomec (R), Merial) and moxidectin (Cydectin (R), Fort Dodge), administered at the doses recommended by the manufacturers. From April 2002 to May 2004, 25 cattle farms were evaluated. on each farm, steers were divided into treatment and control (not treated) groups based on fecal egg counts (FEC). Between 7 and 10 days after the anthelmintics administration, fecal samples were collected from each animal for post-treatment FEC. Fecal cultures from each group were also prepared for larval identification. After treatment, mean FEC reduction (FECR) in treatment groups (compared with control groups) was assessed on each farm. FECR was lower than 90% on 23 farms after ivermectin treatment. on 19 farms, FECR of 100% was recorded following moxidectin treatment; on the remaining 6, FECR ranged from 90% to 97.2%. After albendazole treatment, FECR was higher than 90% on 20 farms and ranged from 47.4% to 84.6% on other 5. After levamisole treatment, FECR was higher than 90% on 23 farms and equal to 47.4% and 73.7% on other 2 farms. Results indicated the presence of resistant Cooperia spp. and Haemonchus spp., especially to ivermectin; on some farms, resistance to albendazole and levamisole was also observed. (C) 2007 Elsevier B.V. All rights reserved.

S-100 dendritic cells in normal and Dermatobia hominis infested cattle skin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of reduction in egg shedding of gastrointestinal nematodes in cattle following administration of anthelmintics

No intuito de avaliar o tempo necessário para redução dos valores de OPG visando identificar o início de atuação dos anti-helmínticos, cinco grupos de novilhos, naturalmente infectados por nematódeos gastrintestinais foram tratados com moxidectina, ivermectina, fosfato de levamisol e sulfóxido de albendazol. O levamisol promoveu redução no número de ovos de nematódeos eliminados nas fezes (R-OPG) de 97,4% 24 horas após a aplicação, a moxidectina de 98,3% após 36 horas, e o sulfóxido de albendazol de 95,9% após 36 horas. Foi registrada a presença de Cooperia spp. e Haemonchus spp. com resistência a ivermectina. A contagem de OPG realizada aos sete dias pós-tratamento apresentou resultados similares aos obtidos nas contagens realizadas 10 e 14 dias após a aplicação dos anti-helmínticos avaliados, demonstrando que o intervalo adequado entre o tratamento anti-helmíntico e o exame para verificar a redução do OPG pode ser de 7 dias.

Relationship of intestinal histology with the resistance to Trichostrongylus colubriformis infection in three breeds of sheep

Avaliaram-se a associação entre o número de células inflamatórias no intestino delgado e a resistência à infecção por Trichostrongylus colubriformis em ovinos de três raças (Santa Inês, Suffolk e Ile de France), naturalmente infectados. Mastócitos, eosinófilos e leucócitos globulares foram quantificados na mucosa intestinal. A concentração de histamina foi estimada em amostras teciduais do intestino, bem como foi determinado o comprimento de machos e fêmeas de T. colubriformis. A resposta celular foi similar na mucosa intestinal das três raças ovinas (P>0,05). Houve grande variação entre os ovinos em relação aos resultados parasitológicos e celulares, mesmo nos animais de mesma raça. em geral, os animais que apresentaram número menor de células inflamatórias tiveram cargas parasitárias maiores, contagens de ovos por grama de fezes mais altas e exemplares de T. colubriformis maiores. Os resultados indicaram que mastócitos, eosinófilos e leucócitos globulares prejudicaram o estabelecimento, o desenvolvimento e a sobrevivência dos parasitas.

Cytokine gene expression in response to Haemonchus placei infections in Nelore cattle

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Immunological responses and cytokine gene expression analysis to Cooperia punctata infections in resistant and susceptible Nelore cattle

Cellular and humoral immune response, as well as cytokine gene expression, was assessed in Nelore cattle with different degrees of resistance to Cooperia punctata natural infection. One hundred cattle (male, weaned, 11-12 months old), kept together on pasture, were evaluated. Faecal and blood samples were collected for parasitological and immunological assays. Based on nematode faecal egg counts (FEC) and worm burden, the seven most resistant and the eight most susceptible animals were selected. Tissue samples of the small intestine were collected for histological quantification of inflammatory cells and analysis of cytokine gene expression (IL-2, IL-4, IL-8, IL-1 2p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, and MUC- 1) using real-time RT-PCR. Mucus samples were also collected for IgA levels determination. Serum IgG1 mean levels against C. punctata antigens were higher in the resistant group, but significant differences between groups were only observed 14 days after the beginning of the experiment against infective larvae (1-3) and 14 and 84 days against adult antigens. The resistant group also presented higher IgA levels against C. punctata (L3 and adult) antigens with significant difference 14 days after the beginning of the trial (P < 0.05). In the small-intestine mucosa, levels of IgA anti-L3 and anti-adult C. punctata were higher in the resistant group, compared with the susceptible group (P < 0.05). Gene expression of both T(H)2 cytokines (IL-4 and IL-13) in the resistant group and T(H)1 cytokines (IL-2, IL-1 2p35, IFN-gamma and MCP-1) in the susceptible group was up-regulated. Such results suggested that immune response to C. punctata was probably mediated by TH2 cytokines in the resistant group and by T(H)1 cytokines in the susceptible group. (C) 2008 Elsevier B.V. All rights reserved.

Moxidectin-resistant nematodes in cattle in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological, histological, and ultrastructural characterization of degenerating salivary glands in females of the cattle-tick Rhipicephalus (Boophilus) microplus (CANESTRINI, 1887) (Acari : Ixodidae)

The present study describes the morphology and ultrastructure of the salivary glands of semi-engorged females of the southern cattle-tick Rhipicephalus (Boophilus) microplus. The acini that compose these glands, at that specific feeding stage, show cells featuring degenerative process of the salivary glands, such as: vacuolated cytoplasm, condensed chromatin, fragmented nuclei, and presence of apoptotic bodies. In addition, the presence of microorganisms was detected, with morphology typical of protozoa, inside these organs. (c) 2005 Elsevier Ltd. All rights reserved.

Morphological, histological, and ultrastructural studies of the ovary of the cattle-tick Boophilus microplus (Canestrini, 1887) (Acari : Ixodidae)

This study presents the morphology of the ovary, as well as the dynamics of the vitellogenesis process in oocytes of the cattle-tick Boophilus microplus. The ovary of these individuals is of the panoistic type; therefore, it lacks nurse cells. This organ consists of a single tubular structure, continuous, and composed of a lumen delimitated by a wall of small epithelial cells with rounded nuclei. In this tick species, the oocytes were classified into six stages varying from I to VI and according to: cytoplasm appearance and presence of the germ vesicle, yolk granules, and chorion. Oocytes of various sizes and at different developmental stages remain attached to the ovary through a cellular pedicel until completing stage V. Afterwards, they are liberated into the lumen and from there to the exterior. Some oocytes (classified as type VI) showed an atypical appearance indicating that some of the cellular components would be undergoing a degenerative process and/or reabsorption. (c) 2004 Elsevier B.V. All rights reserved.