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.

Sorofrequência de infecção pelo vírus da imunodeficiência felina e vírus da leucemia felina em gatos do município de araçatuba, são paulo

Feline immunodeficiency virus and feline leukemia represent important infectious diseases caused by retroviruses. This study aimed to investigate the prevalence of infection by feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) in cats from the municipality of Araçatuba, São Paulo. Blood samples from 302 cats were collected and tested for the presence of antibodies against feline immunodeficiency virus and antigen of feline leukemia virus by ELISA ® Snap- Combo FIV-FeLV (IDEXX Laboratories). The frequency of FIV positivity was 5.63% (17/302) and of FeLV was 0.33% (1/302). Of the 17 cats infected with FIV, nine (52.94%) were symptomatic. There was a prevalence of FIV infection in males (p 0.0316) and cats aged between one and three years (p 0.0324).

Effect of superstimulatory treatments on the expression of genes related to ovulatory capacity, oocyte competence and embryo development in cattle.

Multiple ovulation (superovulation) and embryo transfer has been used extensively in cattle. In the past decade, superstimulatory treatment protocols that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-time AI (FTAI), have been developed for zebu (Bos indicus) and European (Bos taurus) breeds of cattle. There is evidence that additional stimulus with LH (through the administration of exogenous LH or equine chorionic gonadotrophin (eCG)) on the last day of the superstimulatory treatment protocol, called the 'P-36 protocol' for FTAI, can increase embryo yield compared with conventional protocols that are based on the detection of oestrus. However, inconsistent results with the use of hormones that stimulate LH receptors (LHR) have prompted further studies on the roles of LH and its receptors in ovulatory capacity (acquisition of LHR in granulosa cells), oocyte competence and embryo quality in superstimulated cattle. Recent experiments have shown that superstimulation with FSH increases mRNA expression of LHR and angiotensin AT(2) receptors in granulosa cells of follicles >8 mm in diameter. In addition, FSH decreases mRNA expression of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) in oocytes, but increases the expression of both in cumulus cells, without diminishing the capacity of cumulus-oocyte complexes to generate blastocysts. Although these results indicate that superstimulation with FSH is not detrimental to oocyte competence, supplementary studies are warranted to investigate the effects of superstimulation on embryo quality and viability. In addition, experiments comparing the cellular and/or molecular effects of adding eCG to the P-36 treatment protocol are being conducted to elucidate the effects of superstimulatory protocols on the yield of viable embryos.

Efeitos do extrato de parede de levedura em dieta seca sobre a Microbiologia, ácidos graxos de cadeia curta e redução do odor das fezes de gatos adultos

The aim of this study was to evaluate the effects of yeast cell wall extract (YCW) in dry diet on the fecal microbiota, concentration of short-chain fatty acids (SCFA) and on the odor reduction of cats feces. We used 20 animals of both sexes, randomly assigned to four treatments and five repetitions totaling 20 experimental units: 1) dry commercial diet (control); 2) control + 0.2%, 3) control + 0.4%, and 4) control + 0.6% of YCW in dry matter. Enterobacteriaceae and lactic acid bacteria, fecal concentration of acetic, propionic and butyric acids, ammonia nitrogen and sensory panel were performed. There were no significant differences (p> 0.05) for bacterial counts and the concentration of SCFA and ammonia, but in sensory panel a reduction in the odor of feces could be noted with the use of 0.2% of YCW. We concluded that the addition of up to 0.6% YCW had no effect on the microbiology and the concentration of fatty acids, but there is potential for its use as an additive because of the improvement in the odor of feces. However, further studies are needed to understand the mechanisms of action and the effects of prebiotics for domestic cats.

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.

Síndromes oculares secundárias a infecção pelo Herpesvirus felino-1-Revisão

The feline infectious respiratory disease is the most common diagnosed infection in the veterinary clinic routine, being the Feline Herpesvirus1 the most important causal agent. Once infected, the cat will become a lifetime latent carrier, experiencing episodes of viral reactivation and spontaneous spread especially when there is a stress factor involved. This virus acts in the upper respiratory system and is also associated with eye diseases. The diagnosis is made by viral isolation and treatment protocol is based on a topic antiviral therapy, even though many of them are epiteliotoxic and may progress with intense discomfort in felines.The purpose of this paper is to describe the main ocular manifestations and syndromes seen in cats suffering from feline herpesvirus. Conjunctivitis, epithelial and stromal keratitis, corneal ulceration and indolent ulcers are the main ocular manifestations associated with viral infection, whereas symblepharon, keratoconjunctivitis sicca, proliferative keratitis and corneal sequestration are the main eye syndromes that can be observed in infected animals.

Reproductive biology of the guitarfish Rhinobatos percellens (Chondrichthyes, Rhinobatidae) from the São Paulo Coast, Brazil, western South Atlantic Ocean

The reproductive biology of the guitarfish Rhinobatos percellens was studied from 751 specimens caught by bottom pair trawlers off the coast of São Paulo, Brazil, between c. 24° 00′ S; 45° 15′ W and c. 25° 10′ S; 47° 52′ W, from September 2007 to August 2009. The total length (LT) and total mass (MT) relationship for males and females combined was MT = 1·29E-06 LT 3·15 (r = 0·99, n = 751). The mean LT of sexually mature specimens was 548 mm for males and 583 mm for females. Clasper growth was allometric and showed three distinct phases. Most claspers were calcified in specimens of c. 550 mm LT. The mean diameter of the largest oocyte was 29·8 mm, the mean ovarian fecundity was seven oocytes and ovulation occurred between August and November. Uterine fecundity ranged from two to 13 embryos (mean of five embryos). Larger females had higher litter sizes and larger embryos; the size-at-birth was c. 200 mm LT. The hepato-somatic index oscillated seasonally for males and females; the gonado-somatic index had little variation in males, but varied seasonally in females. The presence of many non-pregnant adult females and of encapsulated eggs during two consecutive seasons suggests a resting period between gestations and the possibility of diapause. © 2012 The Authors. Journal of Fish Biology © 2012 The Fisheries Society of the British Isles.

Gametogenesis and reproductive cycle of Melanorivulus aff. punctatus (Boulenger, 1895) (Cyprinodontiformes, Rivulidae) in Chapada dos Guimarães, Mato Grosso, Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of heat stress on development, quality and survival of Bos indicus and Bos taurus embryos produced in vitro

Heat stress is an important cause of poor development and low survival rates in bovine embryos. Experiments were conducted to test the hypothesis that Bos indicus embryos are more resistant to heat stress than are Bos taurus embryos. In experiment 1, Nelore and Jersey embryos from oocyte pick-up-derived oocytes were submitted to heat stress (96 hours post-insemination, 41 °C, 6 hours), developmental ratios were assessed at Day 7 (Day 0 = day of fertilization), and blastocysts were frozen for RNA extraction. Experiment 2 evaluated expression of COX2, CDX2, HSF1, and PLAC8 in previously frozen blastocysts. In experiment 3, Nellore and Angus embryos from oocyte pick-up-derived oocytes were submitted to heat stress (96 hours post-insemination, 41 °C, 12 hours) and transferred to recipients on Day 7. In experiment 4, embryos developed as in experiment 3 were fixed for Terminal deoxynucleotidyl transferase dUTP nick end labeling labeling and total cell counting. In experiment 1, heat stress decreased the percentage of Jersey oocytes that became blastocysts, but had no effect on Nellore embryos (34.6%, 25.0%, 39.5%, and 33.0% for Jersey control, Jersey heat-stressed, Nellore control, and Nellore heat-stressed oocytes, respectively; P < 0.05). In experiment 2, heat stress decreased (P < 0.05) expression of CDX2 and PLAC8, with higher expression of these genes in Nellore embryos than in Jersey embryos. Heat stress also decreased (P < 0.05) expression of COX2 in Jersey embryos, but had no effect on Nellore embryos. Expression of HSF1 was decreased (P < 0.05) by heat stress in both breeds, with a greater effect in Nellore embryos. In experiment 3, heat stress tended (P = 0.1) to decrease the percentage of pregnancies among cows (Day 30 to 35) that received Angus embryos. In experiment 4, heat stress increased (P < 0.05) the percentage of apoptotic blastomeres, but had no breed-specific effects. In addition, Nellore embryos had fewer (P < 0.05) Terminal deoxynucleotidyl transferase dUTP nick end labeling- positive blastomeres than did Angus embryos. We concluded that the detrimental effects of heat stress were dependent upon embryo breed and were more evident in Bos taurus embryos than in Bos indicus embryos. © 2013 Elsevier Inc.

Effect of ricinoleic acid esters from castor oil (Ricinus communis) on the oocyte yolk components of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae)

Rhipicephalus sanguineus are bloodsucking ectoparasites, whose main host is the domestic dog, thus being present in urban areas and closely located to people. Eventually, this tick species parasitize humans and can become a potential vector of infectious diseases. Methods to control this type of pest have been the focus of many research groups worldwide. The use of natural products is increasingly considered nowadays, due to the low toxicity levels to the host and low waste generation to the environment. This study tested the effect of ricinoleic acid esters from castor oil (as an potential acaricide) on the reproductive system of R sanguineus females, more specifically on the vitellogenesis process. For this, two groups were established: the control group (CG) and the treatment group (TG) with five rabbits in each (New Zealand White), used as hosts. NaCl and ester were added to rabbits' food and offered to the hosts. After full engorgement, the females were collected and had their ovaries extracted. The ticks ovaries were submitted to histochemical techniques so the effects of esters could be observed over polysaccharides, proteins and lipids yolk. Changes in the deposition of yolk components were observed. This caused modifications on elements of polysaccharide origin and on glycoprotein compounds, interfering in the final yolk synthesis and compromising the development of the future embryo. © 2012.

Rhipicephalus sanguineus (Acari: Ixodidae) female ticks exposed to castor oil (Ricinus communis): An ultrastructural overview

Tick control has been accomplished through the use of synthetic acaricides, which has created resistant individuals, as well as contaminating the environment and nontarget organisms. Substances of plant origin, such as oils and extracts of eucalyptus and neem leaves, have been researched as an alternative to replace the synthetic acaricides. Ricinoleic acid esters from castor oil have recently been shown as a promising alternative in eliminating bacterial contamination during ethanol fermentation, by acting as an effective biocide. The same positive results have been observed when these esters are added to the food given to tick-infested rabbits. This study tested the effect of these substance on the reproductive system of Rhipicephalus sanguineus females, added to rabbit food, more specifically on oogenesis. For this, four groups were established: four control groups (CG1, CG2, CG3, and CG4) and four treatment groups (TG1, TG2, TG3, and TG4) with one rabbit in each (New Zealand White), used as hosts. After full 4 days feeding (semi-engorgement), the females were collected and had their ovaries extracted. In this study, it was observed that R. sanguineus females exposed to esters had their ovaries modified, which was demonstrated through transmission electron microscopy techniques. The addition of ricinoleic esters to the diet of tick-infested rabbits revealed how toxic such substances are for the cytoplasmic organelles of oocytes and pedicel cells. These compounds can change the morphophysiology of germ and somatic cells, consequently influencing their viability and, therefore, confirming that the ricinoleic acid esters from castor oil are a promising substance in the control of R. sanguineus. © 2012 Springer-Verlag Berlin Heidelberg.

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.

Female germ cell renewal during the annual reproductive cycle in Ostariophysians fish

The objective was to characterize female germ cell renewal during the annual reproductive cycle in two species of ostariophysian fish with distinct reproductive strategies: a siluriform, Pimelodus maculatus, in which oocyte development is group synchronous and the annual reproductive period is short; and a characiform, Serrasalmus maculatus, with asynchronous oocyte development and a prolonged reproductive period. These reproductive strategies result in fish determinate and indeterminate fecundity, respectively. Annual reproductive phases were determined by biometric and histologic analysis of gonads and interpreted according to new proposals for phase classification and stages of oocyte development (with special attention to germinal epithelium activity). Histologically, there were two types of oogonia in the germinal epithelium: single oogonia and those in mitotic proliferation. Oogonial proliferation and their entry into meiosis resulted in formation of cell nests (clusters of cells in the ovarian lamellae). Morphometric analysis was used to estimate germ cell renewal. Based on numbers of single oogonia in the lamellar epithelium, and nests with proliferating oogonia or early prophase oocytes throughout the annual reproductive cycle, oogonial proliferation and entrance into meiosis were more intense during the regenerating phase and developing phase, but decreased sharply (P < 0.05) during the spawning-capable phase. Oogonial proliferation gradually recovered during the regressing phase. We concluded that, independent of species or features of the reproductive cycle, germ cell renewal occurred during the regenerating phase, ensuring availability of eggs for the spawning event. © 2013 Elsevier Inc.

Physiology and endocrinology symposium: Influence of cattle genotype (Bos indicus vs. Bos taurus) on oocyte and preimplantation embryo resistance to increased temperature

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

High incidence of 'Dag-like' sperm defect in the domestic cat

The occurrence of a high incidence of sperm tail defects in a male domestic cat resembling the known 'Dag-like' defect is reported. Sperm analyses were performed in ejaculated samples collected by an artificial vagina and in testicular and epididymal sperm cells after castration. The following alterations were observed using transmission electron microscope: heavily coiled sperm tails containing several axonemal units enclosed in the same common cell membrane; aberrations in the axonemal main structure; and swollen and unevenly distributed mitochondria in the midpiece. Abnormal modifications in the mitochondrial sheath were also found in sperm cells retrieved from testes and epididymides. Considering these findings, we can conclude that this is the Dag-like defect, described previously in other domestic species and a testicular origin may be involved. © ISFM and AAFP 2012.