Reproductive technology in domestic carnivorous

Background: The delay in development of artificial reproduction techniques on carnivorous could be due to countless reasons, but the lack of commercial interest is probably the most important one. The majority of canines are small structures, canidae are extremely fertile and a great number of species are adapted to domestication or captivity. Finally, the canine gamete physiology presents a difficult adaptation of technology knowledge obtained from other species. Furthermore, domestic felines are animals of company and there is no interest in reproducing them in a large scale, as it has been observed in other domestic animals, however, besides of being a valuable model for the development of in vitro techniques, the domestic cat is also used as an embryo receptor for different species of small wild felines due to physiological similarities among them, in vitro embrionary development, Review: It was reviewed the main insights about the reproductive physiology in female dogs, in vitro oocytary maturation (IVM), pregnancy and conception rate with dogs' frozen/unfrozen semen and PIV in domestic cats. The majority of mammal oocytes restart meiosis spontaneously after ovulation and reaches MII in artificial environment; in an in vitro maturation system in bovines, around 90% of oocytes complete their maturation, although its development capacity can be reduced subsequently. The success of IVM in canidae have been limited, with maturation rate varying from 0 to 58%, usually around 20%. The greatest difficulties include oocyte quality, hormonal environment, protein supplementation, cumulus / oocyte cell interaction, donor breed and age, culture systems, oxygen tension, amino acids, growth factor and sequential means. The freezing process reduces the quality of the semen, firstly because it reduces the number of living sperms and secondly because freezing produces cell modifications that could alter the sperm motility, longevity, integrity of membranes and its fertilizing capacity. Conclusion: Nowadays, several researches are being performed with the aim of increasing viability after dogs' and cats' semen is unfrozen, using extenders, cryoprotectors, freezing and unfreezing curves, addition of antioxidant substances. The aim of this text is to inform about the improvements obtained on the artificial reproduction techniques, emphasizing the oocytary maturation in female dogs, semen cryopreservation and artificial insemination in domestic dogs and cats.

Bactérias e resposta inflamatória no útero de cadelas, após inseminação artificial

The present study aimed to obtain information about the uterine inflammatory response (number of polymorphonuclear neutrophilic granulocytes - PMNs) in bitches after artificial insemination (AI) and identify the uterine microflora present after the following treatments: insemination using semen with extender (n=6), insemination with fresh semen (n=6) and no inseminated (n=6). The percentage of PMNs on the endometrial surface and within histological sections was evaluated together with the presence of aerobic bacteria in the uterine lumen. For endometrial cytology, there was no significative difference on the number of inflammatory cells between bitches not inseminated (3.05 ± 1.74 PMNs) and those inseminated with fresh semen (3.55 ± 1.51 PMNs); There was a significative difference in both groups compared to the inseminated with semen plus extender (7.80 ± 1.67 PMNs) (p<0.05). Histology showed that there was no significative difference on the number of inflammatory cells between bitches not inseminated (87.72 ± 35.2 PMNs) and those inseminated with fresh semen (122.97 ± 43.31 PMNs); however, it was observed differences in both groups compared to those inseminated with semen plus extender (171.94 ± 42.74 PMNs) (p<0.05). Eight animals, randomly distributed in the groups, showed the presence of Staphylococcus sp and Proteus sp., in the microbiological exam. The extender for semen, with Tris, is a potent inducer of uterine inflammation, and positive uterine cultures may be obtained during estrus without inflammation or uterine infection.

Influência da raça do touro (Bos indicus x Bos taurus) na tolerância ao estresse térmico calórico de embriões bovinos produzidos in vitro

To better understand the differences related to HS resistance between Bos indicus and Bos taurus, we aim to verify if the HS tolerance is due mostly to the genetic contribution from the oocyte, spermatozoa or both. Oocytes from Nelore and crossbreed Holstein cows (cHST) were collected, matured and fertilized with semen from Nelore (N), Angus (An), Brahman (Bra) and Gir (Gir) bulls. Nine six hours post insemination (hpi), ≥ 16 cells embryos were separated in two groups: control and HS. In control group, embryos were cultured at 39°C, whereas in the HS group, embryos were subjected to 41°C for 12 h, and then returned to 39°C. There was no effect of HS on blastocyst and hatched blastocyst rates in all breeds analyzed. The percentage of oocytes that cleaved and reached morula stage was significantly lower (p < 0.05) in cHST x Gir as compared to the other breeds. Additionally, blastocyst rates was higher in cHST x N than in cHST x An and cHST x Gir (p < 0.05). It was concluded that the oocyte is more important than the spermatozoa for the development of thermotolerance, since the breed of the bull did not influence embryo development after HS.

Análise espermática de perdizes (Rhynchotus rufescens) criadas em cativeiro e suplementadas com selênio

Due to the commercial importance of the red-winged tinamou (Rhynchotus rufescens), for the past few years, the employment of reproductive biotechnologies has been attempted. Thirty animals were randomly assigned into two groups: control group (no selenium) and treatment group (supplemented with 0,2 a 0,8 mg selenium/ 1000 kg ration). Animals were allocated at the FCAV - UNESP/Jaboticabal (2007-2008). Semen collections were performed by digital manipulation and divided in pools of at least 150 μL. After the immediate evaluation of motility, vigour, concentration and morphology, an aliquot of 20 μL was diluted in 300 μL of physiologic solution in order to test acrosome and membrane integrities, which were performed by counting 200 cells for each test. Cells were evaluated as follows: 1) Intact acrosome: lilac acrosome; Non-intact acrosome: pink acrosome; 2) Live cells: non stained; Dead: stained. Data was statistically analysed using the SAS System for Windows. No differences were found between treatment and control groups for volume, motility, vigour, mean number of spermatozoa per animal, concentration, Intact acrosome, Intact membrane. The difference found on midpiece sperm defect (Se = 1,33 ± 0,53 and control = 3,78 ± 0,69, p = 0.0107) may be due to the damages caused by the selenium deficiency to the architecture of the midpiece, which compromises sperm mobility and fertilization capacity.

Composição mineral da carcaça e dos cortes da carcaça de ovinos jovens e adultos

Aiming to evaluate the mineral composition (iron, calcium and sodium) of the carcass and carcass cuts of meat from different ovine categories, 18 1/2 Ile de France 1/2 Polwarth sheeps (6 uncastrated lambs, 6 discarded ewes and 6 discarded wethers) were used. Animals were in grazing system based on Tifton - 85 pastures and were supplemented with concentrate. Lambs were slaughtered at 32 kg of live weight, at around 5 months of age. Ewes and wethers were slaughtered with 55 kg and 60 months of age. The iron values were differents between categories for those valued cuts muscles, with higher average to legs muscles from adults ovine of 3.86 mg/100g with regard to lambs of 1.91 mg/100g, respectively. The iron values of the muscles from half carcass didn't differ for that animal category, with 3.38 mg/100g average. The calcium values in muscles from half carcass and leg cut were higher to wethers (9.84 e 7.35 mg/100g), intermediaries to ewes (6.28 e 6.54 mg/100g) and smallers values to lambs (5.30 e 4.22 mg/100g). There wasn't differences for sodium values to valued categories and muscles, with values around 56.75 mg/100g. In conclusion, the sodium values are similar between animals categories and from differents carcass muscles and that the iron and calcium values present differences.

Produção de forragem de Tifton 85 adubado com nitrogênio e submetido à lotação contínua

The present assay was conducted at the Research and Development of Adamantina, SP, of the Agência Paulista de Tecnologia dos Agronegócios (APTA) - Pólo da Alta Paulista to evaluate the effect of nitrogen fertilization on morphological composition, density of tillers, leaf area index, light interception and herbage accumulation in grazed Tifton 85 subjected to continuous stocking system. The treatments consisted of four levels of nitrogen (0; 100; 200 and 400kg/ha/year of N), arranged in a randomized complete blocks experimental design with four replications. The grasslands of Cynodon cv. Tifton 85 studied influenced the time of evaluation and of nitrogen levels with a marked effect in morphological composition of the pasture, leaf area index, light interception and population density of tillers that were instrumental in the accumulation of forage. It was found a positive linear effect of nitrogen fertilization on the density of vegetative tillers, totals and herbage accumulation nitrogen fertilization had a marked effect on the amount of total and vegetative tillers, leaf area index in light interception and herbage accumulation. The cultivar Tifton 85 has not reached the necessary conditions for crop growth rate maximum when managed under continuous stocking keeping the grass to 10cm tall.

Idade masculina: Qualidade do sêmen e fertilidade: Idade masculina X qualidade seminal

The man's age seems to affect reproductive function but to a lower degree compared with the woman's age. In fact, the effect of paternal age on semen quality has been discussed, but the results are contradictory. Considering the differences in the results of studies, this review aimed to analyze the literature regarding the effect of paternal age on semen parameters (volume, concentration, motility and morphology) in sperm DnA damage in assisted reproduction results and genetic diseases.

Congelação de espermatozoides epididimários de gatos utilizando o diluidor Botu-crio® após refrigeração por 24 h em contêiner de transporte de sêmen Botu-tainer®

The possibilities of using the sperm collected from the epididymis have been widely used because the fertilizing capacity sperm preservation and the possibility of using it for wild cats. But in the process of cryopreservation, some studies show a decrease in the quality of the sperm when left under cooling before frozen for some time. This study aimed to assess the quality of the epididymal sperm obtained from domestic cats after cryopreservation using a diluent based on egg yolk and glycerol (Botu-crio®), comparing the morphofunctional characteristics after cooling for 24 hours in a container of semen transport (Botu-tainer®). We use eight cats submitted to elective orchiectomy, aging from eight months, without racial determination, and good nutritional status. These sperm characteristics were: motility, vigor, concentration, membrane integrity and morphology. It has been found, after statistical analysis, that the container of semen was able to maintain sperm viability, even for 24h. We also observed a significant decrease on all parameters after frozen, consequential, probably to thermal stress that occurs in processing. However, the percentage of membrane integrity after thawing shows good employability of the Botu-crio®, which viability is possible to perform in vitro fertilization, requiring higher ratings.

Possible mechanism by which zinc protects the testicular function of rats exposed to cigarette smoke

Background: The aim of this study was to evaluate the changes in testicular function of rats due to cigarette smoke exposure and the possible mechanism by which zinc protects against these alterations. Methods: MaleWistar rats (60 days old) were randomly divided into 3 groups: control (G1, n = 10); exposed to cigarette smoke (G2, n = 10; 20 cigarettes/day/9 weeks) and exposed to cigarette smoke and supplemented with zinc (G3, n = 8; 20 cigarettes/day/9 weeks; 20 mg/kg zinc chloride daily for 9 weeks, by gavage). After the treatment period, the animals were euthanized, and materials were collected for analyses. Results: G2 rats showed a reduction in body mass; impaired sperm concentration, motility, morphology and vitality; and increased malonaldehyde and thiol group levels and superoxide dismutase activity as compared to G1. Zinc prevented the reduction of sperm concentration and the excessive increase of lipid peroxidation and induced an increase in plasma testosterone levels, wet weight of testis and thiol group concentration. Conclusions: Exposure to cigarette smoke led to harmful effects on testicular function at least partially due to the exacerbation of oxidative stress. Supplementary zinc had an important modulator/protector effect on certain parameters. The mechanism of zinc protection can be through an increase of SH concentration. Thus, zinc supplementation may be a promising addition to conventional treatments for male infertility related to smoking. Copyright © 2012 by Institute of Pharmacology Polish Academy of Sciences.

Effect of roscovitine and cycloheximide on ultrastructure of sheep oocytes

It is believed the temporary meiosis arrest with roscovitine or cycloheximide may improve the in vitro developmental competence of oocytes in different animal species. However, little is known about the effects of these inhibitors on ultrastructure of ovines cumulus-oocyte complexes (COCs). The aim of this study was to evaluate the progression of cytoplasmic maturation and the ultrastructural changes in sheep COCs exposed to roscovitine or cycloheximide, at acceptable concentrations. COCs were in vitro cultured for 24. h in maturation medium (control group) containing 100 μM roscovitine or 1 μg/mL cycloheximide (treatment groups). After this time, some COCs were cultured for further 22. h in inhibitor-free medium. The ultrastructure organization of COCs was evaluated by transmission electron microscopy before (immature group) and after in vitro culture for 24 and 46. h. As expected, signs of immaturity and maturity were observed in immature and control groups, respectively. In treatment with roscovitine, there were cumulus cells degeneration, swelling of mitochondrias, few cortical granules and many vesicles with electron-dense material. However, in cycloheximide treatment there were not signs of degeneration or cellular senescence. Metabolic units and mitochondrial pleomorphism were found in all experimental groups. These evidences demonstrate that roscovitine promoted irreversible ultrastructural changes while cycloheximide did not affect the cytoplasmic maturation. However, the implications on embryo development are still unclear. © 2012 Elsevier B.V.

Rough virulent strain of Brucella ovis induces pro- and anti-inflammatory cytokines in reproductive tissues in experimentally infected rams

The ovine brucellosis caused by Brucella ovis has tropism for reproductive tissues but until now the mechanism of bacterial persistence is not understood. Cytokine expression profiles were studied for 8 months in rams after being experimentally infected with the rough virulent strain of B. ovis (R- B. ovis) to study the pathogenesis of B. ovis and immune mechanism possibly associated to bacteria tropism and persistence. The messenger RNA (mRNA) expression levels of interleukin-1α (IL-1α), IL-1β, IL-6, IL-10, IL-12, interferon-γ (INF-γ) and tumour necrosis factor-α (TNF-α) cytokines were quantified by real-time quantitative RT-PCR (qRT-PCR) in reproductive tissues (epididymus, testicles, ampolae, vesicular glands and bulbourethral glands), and non-reproductive (liver, spleen and kidneys) tissues at 30, 60, 120 and 240 days post infection (dpi). During the acute phase of infection at 30. dpi, the host immune response was most notable demonstrating an up-regulation of several cytokines in reproductive tissues, including the epididymus (IL-6, IL-1β and IL-1α), testicles (INF-γ and IL-12), bulbourethral glands (IL-6 and TNF-α) and ampolae (INF-γ, IL-10, IL-1β and IL-1α). During the development of infection, cytokine gene expression levels decreased, providing evidence of immunosuppression and evidence of immune evasion that favoured persistence of chronic R- B. ovis infection. During the chronic phase of R- B. ovis infection (120 and 240. dpi), cytokine production was down-regulated in the epididymus (IL-1β and IL-1α), testicles (INF-γ and IL-12), and ampolae (INF-γ, IL-10, IL-1β and IL-1α), with the exception of the bulbourethral glands (IL-6 and TNF-α) and epididymus (IL-6); in these tissues, R- B. ovis infection resulted in up-regulation of the pro-inflammatory cytokine IL-6. Herein, we report cytokine expression profiles in tissues of rams experimentally infected with the rough strain of B. ovis, which are associated with bacterial persistence and macrophage activation. © 2012 Elsevier B.V.

Improvement of Matrinxã, Brycon amazonicus, Larviculture by Exposing Eggs to Triiodothyronine

This work evaluated the effect of triiodothyronine (T3) on larviculture of matrinxã, Brycon amazonicus. Oocytes of three females were pooled, fertilized with pooled semen of two males and separated in four batches that were immersed in triiodothyronine solutions as follows: M1 (control - water); M2 (0.01mg/L T3); M3 (0.05mg/L T3); and M4 (0.1mg/L T3). Triiodothyronine did not affect fertilization rate and number of hatched larvae. Weight of hatched larvae was significantly higher in treatments M3 and M4, as well as among larvae sampled at Day 12 in all treatments. After 12d of rearing, biomass gain was higher in the hormone treatments (M1 688±569mg; M2 2436±562mg; M3 3572±569mg; and M4 4129±770mg). In general, coefficients of variation of weight (CVw) and length (CVl) did not differ among treatments and cannibalism was registered between 36 and 72hours post-hatching (h.p.h.) without differences among treatments. Larval survival increased in the hormone treatments (M1 26.5%; M2 37.6%; M3 40.6%; and M4 40.8%). The results indicate that the immersion of matrinxã eggs in triiodothyronine can promote beneficial effects to its larviculture and indicate promising perspectives for culture of this tropical species. © by the World Aquaculture Society 2013.

Prevalência e fatores de risco associados à toxoplasmose ovina no Nordeste do Brasil

In this study, we aimed to determine the prevalence and risk factors associated to Toxoplasma gondii infection in sheep from northeastern Brazil. A total of 932 ovine serum samples from 54 properties in 19 municipalities of the state of Sergipe were collected and assayed using indirect fluorescent antibody test. The assay used antibodies against Toxoplasma gondii (IFAT-IgG) with a cutoff point of 1:64. We observed that 28.22% (263/932) of the ovine samples were serum-reactive. In a logistic regression, factors such as consumption of water directly from the source, consumption of water from a deep well, and age below 12 months were associated with protection; whereas factors such as presence of cats on the property, presence of slatted floor, and use of exchanged or borrowed breeding males were associated with infection. The studied area can be considered endemic for toxoplasmosis, so it is necessary to adopt preventive and control measures because this zoonotic infection poses risks to public health.

Brucella ovis: InvasãO, tráfego, fatores de virulência e resposta imune

Brucellosis remains an economic problem in animals and public health. Worldwide ovine brucellosis caused by Brucella ovis is considered a major cause of infertility in sheep. The factors responsible for persistence of the agent are not known, as well as the mechanisms involved in immune defense and possibly the persistence of the agent. Brucella spp. induces moderate inflammatory response. The nature of the intracellular agent stimulates immune response of the type 1 helper T lymphocytes. Studies of the pathogenesis of ovine brucellosis are scarce. Recent developments have shown that the inflammatory response induced by moderate brucelas represent probably the result of an attempt to escape the immune response and suppression of host immune response. Were reviewed by the mechanisms described by brucelas and Brucella ovis for penetration into the host, escape of the immune response and the immune response generated by the infection.

Proteinograma do soro lácteo de ovelhas da raça santa inês em diferentes fases de lactação

This study aimed to evaluate the influence of lactation phases on the proteinogram of whey protein in Santa Inês ewes. Ewes were accompanied in a semi-intensive system using the same sanitary and nutritional management evaluated at 15, 30, 60 and 90 days postpartum (end of weaning and lactation). Clinical examination of the mammary gland was carried out through and bacteriological culture. The screening of the material resulted in 44 milk samples of healthy glands concurrent negative by CMT and bacteriological culture exam. For obtaining the whey protein renin solution was used. The whey was fractionated into aliquots and kept in the -80C freezer to later separation of protein fractions. For determination of total protein of whey protein was employed the biuret, observing the linearity of the test. Separation of protein fractions was performed, using polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). Eigth protein were observed including lactoferrin, serum albumin, IgA, IgG (heavy chain IgG (IgG CP), light chain IgG (IgG CL), ß-lactoglobulin, a-lactalbumin and proteins identified as PM 15000 and PM 29000. No significant difference was observed at different stages of lactation in the following protein: IgA (P>0.3895), lactoferrin (P>0.1611), PM 29000 (P>0.4879), α-lactalbumin (P>0.0799) and PM15000 (P>0.4494). In total protein (P<0.0022), albumin protein (P<0.0377) and IgG (P<0.0354) it was observed a significant variation in the first moments of observations, in the ß-lactoglobulin protein (P<0.0005) there was significant variation with reduction of 15 to 30 days postpartum with progressive elevation until the last stage of lactation (90 days postpartum). The SDS-PAGE technique allowed the quantification of eigth whey proteins in health ewes. The protein fractions identified reflect the profile of whey to ovine species, with influence of stages of lactation in albumin, IgG and ß-lactoglobulin.