Polymorphonuclear leukocytes CH138+apoptosis evaluation in milk with high and low somatic cell count - preliminary data

Polymorphonuclear leukocyte (PMNL) apoptosis is central to the successful resolution of inflammation. Since Somatic Cell Count (SCC) is an indicator of the mammary gland's immune status, this study sought to clarify the influence that these factors have on each other and on the evolution of the inflammatory process. Milk samples were stained with annexin-V, propidium iodide (PI), primary antibody anti-CH138A. Negative correlation between SCC and PMNL apoptosis was found, and a statistical difference between high SCC group and low SCC group was observed concerning the rate of viable PMNL, apoptotic PMNL, necrotic PMNL and necrotic and/or apoptotic PMNL. Overall, the high cellularity group presented lower proportions of CH138+ cells undergoing apoptosis and higher proportions of viable and necrotic CH138+ cells. Thus, it can be concluded that PMNL apoptosis and SCC are related factors, and that in high SCC, milk apoptosis is delayed. Although there is a greater amount of active phagocytes in this situation, apoptosis' anti-inflammatory effects are decreased, while necrosis' pro-inflammatory effects are increased, which can contribute to chronic inflammation.

Jatropha curcas pericarp toxicity in sheep

Physic nut (Jatropha curcas) is a plant cultivated for biofuel production. Pericarp is a potential livestock food source by-product. However, its use may be limited due to the presence of toxic compounds, mainly phorbol esters. Thus, this study aimed to evaluate pericarp toxicity. Twenty sheep were divided in four groups, one control group which did not receive the plant and three experimental groups which received pericarp in 15% (G15), 30% (G30) and 45% (G45) concentrations for 23 days. After 10 days of treatment, pericarp ingestion produced food intake decrease, diarrhea, dehydration and loss of body condition. All treated groups showed decrease in alkaline phosphatase activity. G30 animals presented reductions in urea and total protein concentrations, and increase in potassium and sodium levels. G45 animals showed increase in serum aspartate aminotransferase activity and in albumin, creatinin, total and indirect bilirubin levels. Anatomohistopathologic findings included ascites, hydropericardium, congestion of the gastintestinal tract and lungs, pulmonary edema and adhesions in the thoracic cavity, renal tubular cells and centrilobular cytoplasmic vacuolation and lymphohistiocytic pneumonia and lymphoplasmacytic and histiocytic enteritis. On the physiochemical analysis 0.3845mg of phorbol esters/g of pericarp were detected. It is concluded that J. curcas pericarp is toxic and is not recommended for sheep feeding.

Genetic correlations and heritability estimates of reproductive traits of Nellore bulls

Estimates of phenotypic, genetics and residual variances for reproductive traits in 5903 Nellore bulls were obtained. The experimental model used was multiple trait derivative-free restricted maximum likelihood. The values obtained for heritability were 0.24 +/- 0.05 for scrotal circumference at 450 days of age and 0.37 +/- 0.05 at 21 months for age at the time of the breeding soundness evaluation; 0.24 +/- 0.05 and 0.26 +/- 0.05 for left and right testicle length; 0.29 +/- 0.05 and 0.31 +/- 0.05 for left and right testicle width; 0.12 +/- 0.04 for testicle format; 0.33 +/- 0.06 for testicle volume; 0.11 +/- 0.03 for gross motility; 0.08 +/- 0.03 for individual motility and 0.05 +/- 0.02 for spermatic vigor; 0.20 +/- 0.04, 0.03 +/- 0.02 and 0.19 +/- 0.04 for larger defects, smaller defects and total defects, respectively. The values for heritability for testicular biometric characteristics were moderate to high while the seminal characteristics, presented low values. Genetic correlations between scrotal circumference with all the reproductive traits were favorable, suggesting the scrotal circumference as a feature of choice in the selection of bulls.

Macroscopic description of teeth of Azara's agouti (Dasyprocta azarae)

Oliveira F.S., Martins L.L., Canola J.C., Oliveira P.T., Pecora J.D. & Pauloni A.P. 2012. Macroscopic description of teeth of Azara's agouti (Dasyprocta azarae). Pesquisa Veterinaria Brasileira 32(1):93-95. Departamento de Medicina Veterinaria, Universidade Estadual de Maringa, Campus Umuarama, Cx. Postal 65, Umuarama, PR 87501-970, Brazil. E-mail: singaretti@ig.com.br The teeth of Azara's agouti (Dasyprocta azarae) were described macroscopically in order to provide biological data on one of the largest wild rodents of the Americas. Radiography was taken on six heads and the teeth were described. Enamel surrounds the coronal dentin, projects to the roots and is present as parallel inner laminae in buccolingual direction. The dentin is located among the enamel laminae and surrounds the pulp horns. The cementum is located internally to the enamel laminae. On the lingual surface, the cementum and dentin are the outer elements.

Genotyping of polymorphisms in the prnp gene in Santa Ines sheep in the State of Sao Paulo, Brazil

Santos C.R., Mori E., Leao D.A. & Maiorka P.C. 2012. [Genotyping of polymorphisms in the prnp gene in Santa Ines sheep in the State of Sao Paulo, Brazil.] Genotipagem de polimorfismos no gene prnp em ovinos Santa Ines no Estado de Sao Paulo. Pesquisa Veterinaria Brasileira 32(3):221-226. Laboratorio de Neuropatologia Experimental e Comparada, Departamento de Patologia, Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Cidade Universitaria, Sao Paulo, SP 05508-270, Brazil. E-mail: caio-patologia@usp.br Enzootic paraplexia or scrapie is a fatal neurodegenerative disease affecting mainly sheep and rarely goats. The disease is influenced by polymorphisms at codons 136, 154 and 171 of prnp gene that encodes the prion protein. The animals may be susceptible or resistant to the development of the disease according to the allelic sequences observed in these codons. In Brazil there were only cases of scrapie in imported animals, therefore the country is considered free of the disease. This study performed the genotyping of different polymorphisms associated to the development of scrapie. Then, based on these findings the animals were categorized in resistant and susceptible. A total of 118 samples were sequenced from the Santa Ines sheep raised on properties located in the State of Sao Paulo. From these samples, 6 alleles and 11 genotypes were identified (ARQ / ARQ, ARR / ARQ, ARQ / AHQ, ARQ / VRQ, AHQ / AHQ, ARR / ARR, ARR / AHQ, VRQ / VRQ, ARQ / TRQ, TRR / TRR, TRQ / TRQ), the genotype ARQ / ARQ presented a frequency of 56.7%. It was also detected the presence of tyrosine at codon 136, which may be considered a rare observation, since there is no report regarding Santa Ines breeding presenting this polymorphism. These results showed the great genetic variability in Santa Ines in Sao Paulo and only 1,69% of the genotypes observed are extremely resistant to scrapie. These data demonstrate that the Santa Ines sheep can be considered potentially susceptible to scrapie.

Quality and quantity of milk produced by Saanen goats submitted to stress during three sequential days

The experiment aimed to study the effect of physiological stress on cortisol levels, quality and quantity of milk through punctual administration of ACTH. Twelve Saanen goats were divided in two experimental groups: ACTH group (0,5 mu g of ACTH/Kg.L.W); Placebo group (placebo solution). Milk production, and percentages of protein, fat, lactose and SCC (somatic cells counting) of the milk were analyzed before, during and after the administration of ACTH/placebo. Simultaneously to the ACTH/placebo administration and during three sequential days, blood was collected to evaluate cortisol concentrations. At times -30 and zero, both groups presented basal concentrations of cortisol. The increase of cortisol contents was significant at times 60 (group ACTH: 59.00 +/- 5.70 and groups placebo: 5.23 +/- 1.37ng/mL) and 120 (group ACTH: 47.96 +/- 9.72 and group placebo: 4.38 +/- 1,14ng/mL) since the cortisol content was higher on the ACTH group. The values returned to the basal level at 300 minutes. Concerning milk production, no differences were found between ACTH and placebo groups. Milk, protein, fat, lactose and SCC did not distinguish one group from another. The results indicated that the physiological stress induced during three days was not harmful to milk production and milk quality of Saanen goats.

Erytrogram of healthy female Holstein calves during the first month of life

Benesi F.J., Teixeira C. M. C., Lisboa J.A.N., Leal M. L. R., Birgel Jr E. H., Bohland E. & Mirandola R. M. S. 2012. [Erytrogram of healthy female Holstein calves during the first month of life.] Eritrograma de bezerras sadias, da raca Holandesa, no primeiro mes de vida. Pesquisa Veterinaria Brasileira 32(4): 357-360. Departamento de Cl nica Medica, Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo, Campus Universitario, Av. Prof. Dr. Orlando Marques de Paiva 87, Sao Paulo, SP 05508-000, Brazil. E-mail: febencli@usp.br In order to establish reference values, and to assess the in fluence of the age factor on the erytrogram of healthy Holstein calves, blood samples from 300 animals were used, spread over 15 experimental groups, according to age, in the first month of life. Variations of the average values for the components of erytrogram were as follows: number of red blood cells (x10(6)/mm(3)) = 6.68-7.60, packed cell volume (%) = 29.80-33.35, hemoglobin concentration (g/dl) = 9.00-10.43, mean corpuscular volume (fl) = 38.93-47.68, mean corpuscular hemoglobin (pg) = 11.75-14.69, mean corpuscular hemoglobin concentration (%) = 29.53-31.63% and number of reticulocytes (x10(3)/mm(3)) = 0.00-11.86. The in fluence of the age factor proved to be significant for the mean corpuscular volume, mean corpuscular hemoglobin and the number of reticulocytes.

Use of Leptospira spp. strains isolated in Brazil in the microscopic agglutination test applied to diagnosis of leptospirosis in cattle herds in eight brazilian states

Sarmento A.M.C., Azevedo S.S., Morais Z.M., Souza G.O., Oliveira F.C.S., Goncales A.P., Miraglia F. & Vasconcellos S.A. 2012. [Use of Leptospira spp. strains isolated in Brazil in the microscopic agglutination test applied to diagnosis of leptospirosis in cattle herds in eight brazilian states.] Emprego de estirpes Leptospira spp. isoladas no Brasil na microtecnica de soroaglutinacao microscopica aplicada ao diagnostico da leptospirose em rebanhos bovinos de oito estados brasileiros. Pesquisa Veterinaria Brasileira 32(7);601-606. Universidade de Sao Paulo, Faculdade de Medicina Veterinaria e Zootecnia, Departamento de Medicina Veterinaria Preventiva e Saude Animal, Av. Prof. Dr. Orlando Marques de Paiva 87, Sao Paulo, SP 05508-270, Brazil. E-mail: savasco@usp.br The aim of this study was to investigate the adequacy of the use of autochthonous strains of leptospires isolated in Brazil, added to antigen collection of the microscopic agglutination test (MAT) applied to the diagnosis of bovine leptospirosis. By means of non-probability sampling, 109 farms and 9,820 cattle, females at reproductive age were chosen from 85 municipalities in the states of Goias, Mato Grosso, Mato Grosso do Sul, Minas Gerais, Parana, Rio Grande do Sul, Santa Catarina and Sao Paulo. Among the 9,820 examined animals, 5,806 (59.12%) were reactants at MAT for at least one serovar using the 23 reference serovars. Employing the collection of reference serovars and the ten autochthonous strains, 6,400 (65.24%) reactants and significant difference (p=0.001) was found. The most probable serovars identified by the collection of reference antigens were Hardjo (43.03%), Shermani (20%), Wolfi (9.96%), Grippothyphosa (5.42%) and Pomona (4.28%). With the collection amplified with the ten strains isolated in Brazil, the most probable serovars were Hardjo (31%), Guaricura-M4/84 (22.50%), Shermani (15.43%), Wolffi (4.76%), Grippothyphosa (3.71%) and Autumnalis (3.24%). The serovar Guaricura, strain M4/84, isolated from bovines and buffaloes in the State of Sao Paulo, was ranked as one of the three most probable serovars in the states of Goias, Mato Grosso, Mato Grosso do Sul, Minas Gerais and Sao Paulo. The addition of autochthonous strains to the MAT antigen collection provided the confirmation of the diagnosis of leptospirosis in 594 cattle (6%) which have been classified as non-reactants by the reference collection (p=0.001).

In vitro differentiation of mesenchimal stem cells of dogs into osteogenic precursors

Lima S.A.F., Wodewotzky T.I., Lima-Neto J.F., Beltrao-Braga P.C.B. & Alvarenga F.C.L. 2012. [In vitro differentiation of mesenchimal stem cells of dogs into osteogenic precursors.] Diferenciacao in vitro de celulas-tronco mesenquimais da medula ossea de caes em precursores osteogenicos. Pesquisa Veterinaria Brasileira 32(5):463-469. Departamento de Reproducao Animal e Radiologia Veterinaria, Faculdade de Medicina Veterinaria e Zootecnia, Universidade Estadual Paulista, Campus de Botucatu, Distrito de Rubiao Junior s/n, Botucatu, SP 18618-970, Brazil. E-mail: silviavet@usp.br The aim of our research was to evaluate the potential for osteogenic differentiation of mesenchimal stem cells (MSC) obtained from dog bone marrow. The MSC were separated using the Ficoll method and cultured under two different conditions: DMEM low glucose or DMEM/F12, both containing L-glutamine, 20% of FBS and antibiotics. MSC markers were tested, confirming CD44+ and CD34- cells with flow cytometry. For osteogenic differentiation, cells were submitted to four different conditions: Group 1, same conditions used for primary cell culture with DMEM supplemented media; Group 2, same conditions of Group 1 plus differentiation inductors Dexametazone, ascorbic acid and beta-glicerolphosphate. Group 3, Cells cultured with supplemented DMEM/F12 media, and Group 4, same conditions as in Group 3 plus differentiation inductors Dexametazone, ascorbic acid and beta-glicerolphosphate. The cellular differentiation was confirmed using alizarin red and imunostaining with SP7/Osterix antibody. We observed by alizarin staining that calcium deposit was more evident in cells cultivated in DMEM/F12. Furthermore, by SP/7Osterix antibody immunostaining we obtained 1:6 positive cells when using DMEM/F12 compared with 1:12 for low-glucose DMEM. Based on our results, we conclude that the medium DMEM/F12 is more efficient for induction of differentiation of mesenchymal stem cells in canine osteogenic progenitors. This effect is probably due to the greater amount of glucose in the medium and the presence of various amino acids.

Morphological aspects of the ovaries of turtle Kinosternon scorpioides raised in captivity

Chaves E.P., Oliveira S.C.R., Araujo L.P.F., Oliveira A.S., Miglino M.A., Abreu-Silva A.L., Melo F.A. & Sousa A.L. 2012. Morphological aspects of the ovaries of turtle Kinosternon scorpioides raised in captivity. Pesquisa Veterinaria Brasileira 32(7):667-671. Departamento das Clinicas, Curso de Medicina Veterinaria, Universidade Estadual do Maranhao, Cidade Universitaria Paulo VI, Tirirical, Sao Luis, MA 65050-150, Brazil. E-mail: alana@elo.com.br The swear turtle "jurara" (Kinosternon scorpioides) is a mud turtle of the Amazon region exposed to disordering capture in the rural areas of Maranhao, Brazil. Despite its popularity in these areas, little meaningful information regarding the reproductive morphology is currently available, fact that impedes the adoption of policies for preservation of the species. To obtain more information, we studied the ovarian morphology adult jurara females kept in captivity by morphological and morphometric analysis in the dry and rainy season. The results revealed that all females were sexually mature and were in a vitellogenic period. The ovaries are two irregular structures composed by follicles in different stages of development (primary, secondary and tertiary) scattered in a stroma of loose connective highly vascularized tissue. The ovary weight was 6.25+/-4.23g and 2.27+/-1.42g, for the right and left one respectively. The gonadosomatic indexes were 2.06% for the dry season and 1.79% for the rainy season. The average of the follicles was 29.83 units per ovary. Microscopically, the mature ovaries revealed a basal layer composed by four cellular layers: the inner and outer theca, stratum granulosum with perivitelline membrane and zona radiata with vitelline membrane. No significant differences were observed in the ovaries either in the dry or wet period.

Validation of a new proposal of quantitative sperm culture for industrialized bull semen

The Brazilian Ministry of Agriculture (MAPA) has discussed the mandatory culture of industrialized semen, both to ensure biosefety, and to prevent in vitro fertilization problems caused by oocyte contamination with ubiquitous and opportunistic bacteria from preputial microbiota. Pour plate, a quantitative technique recommended by the World Organization for Animal Health (OIE), is operationally difficult and costly for routine analysis in Artificial Insemination Centers (AICs). The objectives of this study were to evaluate and validate viable superficial bacteria counts (VSBC), in CFU/mL, compared with pour plate counts, in industrialized bull semen samples from AICs. Semen straws from Projeto Hungria - MAPA bulls were used. VSBC and pour plate were carried out in parallel in serial dilutions of the samples, from 10(-1) to 10(-5). CFU/mL means or medians recorded in each dilution and technique were compared, and no statistical differences were observed between the two techniques regarding the quantification of bacteria in CFU/mL, suggesting that it may be possible to replace pour plate for CBSV, a cheaper and more practical technique.

Germline mutation analysis of Tpit in Poodle dogs with ACTH-dependent hyperadrenocorticism

There is a high incidence of pituitary-dependent hyperadrenocorticism (PDH) in Poodle dogs, with family members being affected by the disease, suggesting a genetic involvement. Tpit is an obligate transcription factor for the expression of pro-opiomelanocortingene and for corticotroph terminal differentiation. The aim of the present study was to screen the Tpit gene for germline mutations in Poodles with PDH. Fifty Poodle dogs (33 female, 8.71 +/- 2.8 years) with PDH and 50 healthy Poodle dogs (32 females, 9.4241 2.8 years) were studied. Genomic DNA was isolated from peripheral blood, amplified by PCR and submitted to automatic sequence. No mutation in the coding region of Tpit was found, whereas the new single nucleotide polymorphism p.S343G, in heterozygous state, was found in the same frequency in both PDH and control groups. We concluded that Tpit gain-of-function mutations are not involved in the etiology of PDH in Poodle dogs.

Lactation stage and udder health status of Santa Ines ewes

FAPESP [2003/08582-7]

Seroprevalence and risk factors associated with Chlamydophila abortus infection in dairy goats in the Northeast of Brazil

Santos C.S.A.B., Piatti R.M., Azevedo S.S., Alves C.J., Higino S.S.S., Silva M.L.C.R., Brasil A.W.L. & Gennari S.M. 2012. Seroprevalence and risk factors associated with Chlamydophila abortus infection in dairy goats in the Northeast of Brazil. Pesquisa Veterinaria Brasileira 32(11):1082-1086. Unidade Academica de Medicina Veterinaria, Centro de Sa de e Tecnologia Rural, Universidade Federal de Campina Grande, Av. Universitaria s/n, Bairro Santa Cecilia, Patos, PB 58700-970, Brazil. E-mail: sergio.azevedo@pq.cnpq.br Few data are available on the prevalence and risk factors of Chlamydophila abortus infection in goats in Brazil. A cross-sectional study was carried out to determine the flock-level prevalence of C. abortus infection in goats from the semiarid region of the Paraiba State, Northeast region of Brazil, as well as to identify risk factors associated with the infection. Flocks were randomly selected and a pre-established number of female goats >= 12 mo old were sampled in each of these flocks. A total of 975 serum samples from 110 flocks were collected, and structured questionnaire focusing on risk factors for C. abortus infection was given to each farmer at the time of blood collection. For the serological diagnosis the complement fixation test (CFT) using C. abortus S26/3 strain as antigen was performed. The flock-level factors for C. abortus prevalence were tested using multivariate logistic regression model. Fifty-five flocks out of 110 presented at least one seropositive animal with an overall prevalence of 50.0% (95%; CI: 40.3%, 59.7%). Ninety-one out of 975 dairy goats examined were seropositive with titers >= 32, resulting in a frequency of 9.3%. Lend buck for breeding (odds ratio = 2.35; 95% CI: 1.04-5.33) and history of abortions (odds ratio = 3.06; 95% CI: 1.37-6.80) were associated with increased flock prevalence.

Demographic characterization of supervised dog and cat populations in Sao Paulo city, Brazil

This study aimed to estimate the owned dog and cat populations of Sao Paulo city using a complex sample with random selection in two stages. In each administrative district, six census sectors and 20 households in each sampled sector were visited from September 2006 to September 2009, totalizing 11,272 interviews. The human: dog ratio was 4.34 and the human: cat ratio was 19.33. The dog population was 2,507,401 and the cat population, 562,965. The dog population was composted of 52.7% males, while among the cat population it was 45.1%. The proportion of sterilized females (23.4% among dogs and 46.1% among cats) was higher than males (11.4% among dogs and 31.5% among cats). The mean age of dogs was 4.99 years, and for cats, 3.53 years. The proportion of restricted (without access to the street) dogs, 64.4%, was higher than restricted cats, 42.5%. The average number of animals/household was 1.60 for dogs and 1.69 for cats. The animal ownership is associated with cultural factors, therefore the characterization of canine and feline population is essential to implement an adequate animal population management program and zoonosis control.