Guilherme Ribeiro Valle

Dogs with Leishmania chagasi infection have semen abnormalities that partially revert during 150 days of Allopurinol and Amphotericin B therapy

The goal of the present study was to characterize the semen quality of dogs naturally infected with Leishmaniachagasi, and treated with Allopurinol and Amphotericin B. Eight naturally infected and eight non-infected dogs were selected. Following semen collection, progressive motility, vigor, concentration and sperm morphology were evaluated. The seminal patterns in the treated animals were evaluated at the beginning (d0) and at days 30 (d30), 60 (d60) and 150 (d150) of treatment. The progressive motility at d0 (35.7+/-22.3%) was less than that of the control group (77.8+/-7.1%) (P<0.05). The vigor was similar to the control group throughout the treatment (P>0.05). The number of sperm/mL, sperm/ejaculate and sperm/kg of body weight was similar among groups (P>0.05). The percentages of normal spermatozoa of infected and treated animals were similar throughout the treatment and to the control group (69.1+/-8.7%) at d60 (37.5+/-11.2%) and d150 (48.3+/-10.8%) (P>0.05), but smaller at d0 (22.7+/-10.5%) and d30 (28.8+/-15.9%) (P<0.05). A greater percentage of acrosome damage was observed in the control group (3.1+/-2.3%) compared to the d60 (0.1+/-0.2%) (P<0.05). The infected dogs had a greater percentage of principal piece defects at d60 (37.0+/-6.3%) than the control group (16.8+/-7.3%) (P<0.05); and greater percentages of detached normal heads at d0 (28.7+/-19.7%) and d30 (18.5+/-18.5%) than the control group (0.4+/-0.5%) (P<0.05). This reduction in semen quality of the infected animals is suggestive of an epididymal dysfunction. Due to this poor semen quality, caution is recommended when using infected male dogs for reproductive purposes.

Aplasia segmentar de corpo uterino em cadela sem raça definida: relato de caso

A case of partial segmental aplasia of the uterine body in a sexually mature mongrel bitch in anestrous phase and normal clinical condition is reported. At necropsy, no changes in the genital organs were observed with the exception of the uterine body that had a segmental (2mm in length) decrease in diameter in its medial-caudal portion, with absence of lumen and hydrometra in the cranial portion of the uterine body and uterine horns. Histopathologic evaluation indicated reduction in thickness of the whole uterine body at the site of stenosis, with all layers preserved including perimetrium, myometrium, and endometrium, but with the absence of a lumen. The walls of the uterine horns were thin and there was a marked decrease in the number of endometrial glands. There were several follicles on different stages of development and corpora lutea in regression, indicating that the bitch had normal cyclic activity.

Detection of Leishmania infantum in the smegma of infected dogs

Considering the venereal transmission of visceral leishmaniasis from dogs to bitches, the aim of this study was to verify if the penile surface and smegma from infected dogs can be the source of parasites in bitches. Twelve Leishmania infantum infected dogs had semen and smegma samples collected for submission to PCR identification of the DNA of the parasite. Semen (41.7%) and smegma (50.0%) have similar positive incidence (P>0.05; Fishers exact test), with 58.3% of the dogs positive for semen and/or smegma samples. The proportion of positivity for both semen and smegma was 33.3%, but 8.3% was positive only for semen, and 16.7% only for smegma, revealing a moderate agreement between tests (K=0.5; Kappa index). It was concluded that Leishmania infantum is present in the smegma of contaminated dogs and it can be a source of parasites for the semen and the bitch.

Efeito da centrifugação e do líquido prostático homólogo na criopreservação de espermatozoides epididimários caninos

Realizaram-se dois experimentos de criopreservacao de espermatozoides epididimarios caninos, investigando-se o efeito da centrifugacao e da adicao do liquido prostatico sobre as caracteristicas fisicas do espermatozoide pos-descongelacao. No experimento I, foi testado o efeito da centrifugacao. As amostras congeladas sem centrifugacao apresentaram pos-descongelacao: motilidade total (MT) de 26,7±21,2%, motilidade progressiva (MP) de 21,2±20,1% e vigor espermatico (V) de 2,2±1,3, e as congeladas apos a centrifugacao: MT de 23,9±17,9%, MP de 20,6±17,4% e V de 2,2±1,0. No teste de termorresistencia, o periodo medio de duracao com MT minima de 10% foi de 165±21,2 minutos sem centrifugacao e de 77,5±63,6 minutos para as centrifugadas, indicando maior longevidade espermatica das amostras nao centrifugadas. No experimento II, foi avaliado o efeito da adicao de liquido prostatico homologo no meio diluidor. As amostras congeladas sem liquido prostatico no meio diluidor apresentaram MT de 13,3±13,1%, MP de 10,9±11,4% e V de 2,1±1,2, e as congeladas com liquido prostatico MT de 14,1±12,6%, MP de 12,2±11,6% e V de 2,2±1,3. Os resultados sugerem que a centrifugacao e a adicao de 10% de liquido prostatico ao diluidor nao tiveram efeito sobre as caracteristicas fisicas do espermatozoide epididimario canino pos-descongelamento.

Sex ratio of equine offspring is affected by the ages of the mare and stallion.

The aim of this study was to determine the influence of parental age on the sex ratio of offspring in horses. Two trials were performed. In the first trial, the data from a randomly obtained population with a 1:1 sex ratio of 59,950 Mangalarga Marchador horses born in Brazil from 1990 to 2011 were analyzed. The sex ratios of the offspring were compared among groups according to the mare and the stallion ages (from 3 to 25 years). In the first step of the analysis, the mares and stallions were grouped according to age in 5-year intervals. In the second step, the groups were based on the parental age gap at conception. In the third step, the group of the mares and stallions with similar ages from the second step was subdivided, and the different parental age subgroups that were divided into 5-year intervals were compared. In the fourth step, the sex ratio of the offspring was determined according to the ages of the mares and the stallions at conception. The second trial was based on the data from 253 horses of several breeds that were born after natural gestation into a herd from 1989 to 2010, and the offspring of groups that were younger or older than 15 years were compared. The data from both trials were analyzed using a chi-square test (P ≤ 0.01 for the first trial; and P ≤ 0.05 for the second trial) for the comparisons of the sex ratios. In the first trial, the Spearman test (P ≤ 0.01) was used to verify the correlations between the parental age and the offspring sex ratio. In the first trial, the offspring sex ratio decreased as the mare or stallion age increased, and the decrease was more marked for the mares than for the stallions. In the second trial, the mares older than 15 years had more fillies than the younger mares, but the stallion age had no effect on the sex of the offspring. The first trial, with a large number of horses, revealed the pattern of the distribution of the sex ratios of offspring according to the parental age in horses, whereas the second trial, with a more restricted number of horses, confirmed the influence of the age of the mare on the offspring sex ratio. We concluded that the parental age affected the offspring sex ratio in horses and that this effect was stronger for the mares than for the stallions.

Detection of Simonsiella species in the vagina of a bitch in heat

THE vaginal bacterial flora of bitches originates from the bowel and from the skin (Van Duijkeren 1992, Johnston and others 2001); a similar microbial flora is found in the prepuce of male dogs (Ling and Ruby 1978). The same microorganisms have been found in healthy bitches and in bitches with vaginitis, cervicitis or metritis, providing evidence that such conditions are caused by opportunistic pathogens (Platt and Simpson 1974, Hirsh and Wiger 1977). The genus Simonsiella is a member of the Neisseriaceae family and is characterised by unique morphology: its cells are wide (1·9 to 6·4 μm), short (0·5 to 1·3 μm), relatively flat (0·5 to 1·3 μm), and attach to each other to form eightto 12cell long, single-series filaments (Hedlund and Staley 2002) that show a transversal striation (Kuhn and others 1978). The ventral surface of Simonsiella species attaches itself to epithelial cells in the host’s oral cavity and upper respiratory tract, and the bacterium is able to move on this surface (Hedlund and Staley 2002). Simonsiella species are Gram-negative (Bruckner and Fahey 1969). Bacteria of the genus Simonsiella reside in the oral cavity, where they are more abundant on the palate than on the gums (Nyby and others 1977). They have been isolated from many species (Bruckner and Fahey 1969, Nyby and others 1977, Kuhn and others 1978, Hedlund and Staley 2002). There have been no reports of the existence of Simonsiella species bacteria in the vagina. This short communication describes the isolation of Simonsiella species from the vagina of a bitch. During a routine examination for the reproductive control of a five-year-old bitch belonging to a privately owned rottweiler kennel (bitch 1) cytological examination of the vagina revealed the presence of unusual microorganisms. Ten days later, fresh samples were collected for cytological examination from the vaginal and oral mucous membranes of the bitch, and also from two other bitches from the same kennel (bitches 2 and 3). Samples were also collected from the preputial and oral mucous membranes of a dog at the same kennel (the sire). The samples were stained with the WrightGiemsa and Gram methods. According to the routine technique for the evaluation of vaginal cytology described by Johnston and others (2001), the first cytological examination of the vaginal sample from bitch 1 showed that the animal was in oestrus; subsequently, it was mated twice with the sire. The slide showed a predominance of superficial cells and a large number of bacteria, as well as some unusual microorganisms. It was suspected that these might be bacteria of the genus Simonsiella. The microorganisms measured on average 13 μm long and 5 μm wide, and had traverse striations and rounded edges. They were present in large numbers over the exfoliated cells (Figs 1, 2). Cytological examinations of the oral mucous membranes of all the animals showed the same unusual microorganisms over the cells. However, 10 days after the first cytological examination of bitch 1, vaginal cytology of all three bitches showed that none of them was in heat, and the unusual microorganisms were no longer present. The organisms were also absent from preputial sample from the sire. Bitch 1 became pregnant and delivered five puppies. The morphology of the microorganisms found in the vaginal and oral samples, and their Gram-negative staining characteristics, were sufficient to classify them as members of the genus Simonsiella (Fig 3). Although a great number of bacteria exist in the vaginal mucous membranes of healthy bitches (Osbaldiston and others 1972, Ling and Ruby 1978, Baba and others 1983, Bjurstrom and Linde-Forsberg 1992, Van Duijkeren 1992, Johnston and others 2001), Simonsiella species had not previously been reported. The Simonsiella species detected in the first cytological examination of the vaginal sample of bitch 1 may have been acquired as a result of its vulva being licked by other dogs or by itself, since the microorganism was present in the oral samples of all the dogs studied, leading to the epithelial surface of its vagina being colonised. It is well known that the number of vaginal bacteria detected by cytological examinations of bitches in oestrus is higher than at any other stage of the canine oestrous cycle (Olson and Mather 1978, Baba and others 1983, Johnston and others 2001), due mainly to the smaller number of neutrophils on the epithelial surface (Johnston and others 2001). Therefore, the Simonsiella species from the oral cavity of bitch 1 or the other dogs may have encountered favourable conditions to attach to and colonise the vaginal mucous membrane. The fact that the Simonsiella species was not detected in vaginal samples from bitches 2 and 3, which were not in heat, or from the second sample taken from bitch 1 after it had been mated and become pregnant, indicates that the microorganism was present only transiently in the vaginal mucous membrane, unlike its behaviour in the oral mucous membrane, which it can colonise (Nyby and others 1977). Veterinary Record (2006) 159, 156-157

Leishmania infantum is present in vaginal secretions of naturally infected bitches at lower levels in oestrogenized bitches than in non-oestrogenized bitches

Naturally Leishmania infantum infected bitches were divided into oestrogenized (n = 11) and non-oestrogenized (n = 6) groups. Vaginal secretions were collected for polymerase chain reaction (PCR), and vulval, vaginal and uterine tissues were collected for the immunohistochemical (IHQ) identification of L. infantum. Parasite DNA was identified in vaginal secretions of non-oestrogenized (41.8%) and oestrogenized (18.2%) bitches (P<0.05; Fisher’s Exact test). IHQ was positive in vulvar dermis (23.5%) and vaginal mucosa (17.7%) but negative in endometrium of all bitches. Poor association between positive vaginal secretion PCR and tissue IHQ (Kappa index) were observed. The results showed that genital secretions are a potential source for dog contamination.

Efeito da radiação vermelha de baixa intensidade sobre o sêmen canino criopreservado

The aim of this study was to analyze the effect of low intensity red light on some kinetic parameters of cryopreserved canine sperm. Ejaculates from 08 adult dogs were centrifuged, diluted in Tris-egg yolk with 6% glycerol, and subsequently separated into: T1: incidence of red light (660 nm) (Fisioled -MMOptics - 100mW) for 60 seconds before the cooling and after thawing, T2: just before cooling, T3: only after thawing, and T4: no incidence. After thawing the samples were subjected to TTR using Sperm analyzer(r). In TTR0, TTR60 and TTR90 there were no differences between the variables analyzed by CASA. Only in TTR30 the effects of the incidence of red light were visible and significant in T1 and T2. T1 resulted in low MT (12.5 ±10.6%) and T2 determined the best result of MT 26.1%±40.3. Similarly T1 showed a higher number of static spermatozoa (77.5±28.9%) compared to T2 (50.6±28%). We concluded that the double incidence of low intensity red light, before cooling and after thawing, had a deleterious effect on canine sperm motility expressed at 30 minutes after thawing.

Identificação imuno-histoquímica de VEGF e IGF-1 em ovários de cadelas no anestro e estro

In order to verify the presence of VEGF and IGF-1 in the ovaries of bitches, immunohistochemical analyzes of the cortical stroma; theca and granulosa of secondary, tertiary and tertiary luteinized preovulatory follicles; and oocytes of primary, secondary and tertiary follicles of ovaries from five bitches in anestrous (Anest) and five in estrus (Est) was performed. The identification of the phases of the estrous cycle was performed by vaginal cytology associated with the measurement of plasma progesterone. The ovaries were treated for immunohistochemical identification of VEGF (PU 360 primary antibody-UP, Biogenex, USA, dilution 1:30) and IGF-1 (primary antibody PabCa, Gro-Pep, Australia; 1:100 dilution). The immunostaining index (MI) was determined for each tissue by the ratio of positively marked area divided by total analyzed area. For oocytes immunostaining was determined as positive or negative. Comparisons of IM between tissues were performed with the Wilcoxon test (deferent tissues in the same group) or Mann-Whitney test (same tissue between different groups), all at 5% significance level. VEGF and IGF-1 have been similarly identified (P>0.05) in all structures evaluated in both groups. It is concluded that in bitches in estrus and anestrous these growth factors are present in ovary cortical stroma, follicles at different stages of development and oocytes.

Taxas de perda gestacional até 60 dias são afetadas por características cíclicas da égua receptora de embrião Mangalarga Marchador

Pregnancy rates were evaluated at 15 days and pregnancy loss between 15 and 60 days on 430 embryo transfers (ET) in Mangalarga Marchador mares. Pregnancy diagnosis was performed between 15 and 60 days after ET. To evaluate the effects of the duration of the follicular phase of the recipient mare, three groups were formed: up to 3 days ( 7d). To evaluate the effects of the size of the pre-ovulatory follicle of the recipient mare, three other groups were formed: below or equal to 35mm (O 45). The groups were compared by Chi-square test (P 7d - 86.0%), and greater pregnancy loss in >7d (24.4%) than in 45 - 81.9%) and pregnancy loss (O 45 - 10.5%) were similar. Reasons for the greatest pregnancy loss in the >7d group have not been elucidated, but we conclude that the duration of the follicular phase may be a factor for choosing recipient mares.