Establishment of a Pregnancy Following Intravaginal Insemination with Epididymal Semen from a Dog Castrated due to Benign Prostatic Hyperplasia

Benign prostatic hyperplasia was diagnosed in an American Staffordshire Terrier of high breeding value presenting concurrent haematuria. Castration as a treatment was synchronized with the oestrus cycle of a bitch selected for insemination. After castration the cauda epididymis was flushed with Gent semen extender and collected spermatozoa were filtered and analysed by Hamilton Thorn computer assisted sperm analysis. A total of 7 ml semen containing 742 x 10(6) spermatozoa with 76.5% mean motility was used for insemination. Intravaginal insemination of the bitch was performed with an insemination catheter for dogs (Kruuse, Marslev, Denmark) on the day when plasma progesterone levels reached 9.9 ng/ml. Normal pregnancy without complications resulted in eight live-born puppies 63 days after insemination. This is the first report of a normal pregnancy and birth of puppies from a bitch inseminated with epididymal semen obtained from a dog affected by benign prostate hyperplasia.     

Subclinical leishmaniasis associated with infertility and chronic prostatitis in a dog

A stud dog was presented for acquired infertility. Haematospermia and teratozoospermia were found on two ejaculates 2 weeks apart. A presumptive diagnosis of prostatitis was made follo-wing ultrasound examination. An ultrasound-guided needle core biopsy was performed under general anaesthesia, revealing a mild chronic macrophagic and plasma cell prostatitis with intracytoplasmic amastigotes consistent with Leishmania spp. infection. Presence of Leishmania infantum, Leishmania donovani or Leishmania chagasi was confirmed by polymerase chain reaction in seminal plasma. Serology and serum protein electrophoresis confirmed the diagnosis of a subclinical active systemic leishmaniasis. A meglumine antimoniate and allopurinol treatment was given which clearly improved within 3 months both general condition and the quality of sperm. To the authors' knowledge, this is the first reported case of a prostatitis secondary to a Leishmania spp. infection. Subclinical systemic leishmaniasis should be considered in the differential diagnosis of infertility in dogs suffering from semen alterations.     

Acute phase protein response in experimental canine leishmaniasis

Acute phase proteins (APPs) have been proposed as useful markers for the diagnosis and monitoring of treatment of dogs infected by Leishmania infantum. However, the kinetics and behavior of these proteins in canine leishmaniasis is still unknown. The aim of this study was to monitor the kinetics of APPs in dogs experimentally infected with L. infantum, before, during and after therapy against canine leishmaniasis. Levels of serum haptoglobin, serum amyloid A and C-reactive protein from 6 infected beagles, positive by both PCR and parasite culture, were monitored for 7 months post-infection. The dogs were then treated for 3 months with allopurinol (20 mg mg/kg/day PO), and their response to therapy was followed for 11 additional months. Levels of Immunoglobulins G and M were recorded during these 21 months and compared. Experimental infection with L. infantum amastigotes induced an increase in all APPs studied which was statistically significant 2 months after infection for all proteins. Clinical recovery was accompanied by a significant decrease of all APPs 1 month after the beginning of treatment. However, differences were found between the APPs in both magnitude and duration of serum level elevations. The increase in total IgG and IgM was delayed in comparison to APPs and contrarily to the APPs, these immunoglobulins did not significantly decrease with treatment. In conclusion, the results of this study suggest that APPs could be used as early markers for disease as well as for monitoring the response to treatment in canine leishmaniasis.     

Perineal hernia because of retroflexion of the urinary bladder in a rottweiler bitch during pregnancy

A young pregnant rottweiler bitch was presented with a large mass of tissue protruding through the vulvar lips. Foetal death was diagnosed on ultrasonography, and ovariohysterectomy was performed, at which time the bladder was observed to be dislocated caudally into the pelvic cavity under the vagina. The bladder was manually repositioned during surgery. One week later, the bitch was healthy with no evidence of dysuria, stranguria or urinary incontinence. Two months later, the owner reported that the bitch was clinically normal with no recurrence of the retroflexion. To the authors' knowledge, this is the first reported case of retroflexion of the urinary bladder during pregnancy.     

Prenatal and transmammary infections of Toxocara canis in dogs: effect of benzimidazole-carbamate anthelmintics on various developmental stages of the parasite

Fenbendazole and albendazole, given at a dose rate of 150 mg/kg for 3 days, produced a 90 per cent reduction in the numbers of second stage larvae of Toxocara canis present in the tissues of dogs although no reduction in the number of larvae found in the brains of infected dogs occurred with this treatment. The results suggest that a course of 3 day therapy with these anthelmintics should prevent prenatal infections in puppies. However, if infection is acquired by bitches during late pregnancy or early lactation, the transmammary route of infection becomes important. Therefore, anthelmintic treatment of the bitch prior to pregnancy will not prevent transmission of infection to her puppies should the bitch acquire a new infection of T. canis during pregnancy or early lactation. Alternatively, infection with T. canis can be controlled through the treatment of neonatal puppies for migrating larvae of T. canis. Treatment of newborn puppies with fenbendazole, albendazole or oxfendazole at a dose of 100 mg/kg for 2–3 days produced a 91–99 per cent reduction in the number of adult parasites found. In addition, a single dose of fenbendazole, given at a dose rate of 40 mg/kg, eliminated 93–96 per cent of adult T. canis from the intestines of 4–5-week-old puppies. These latter treatments would need to be repeated to eliminate completely the infection from puppies.     

Longitudinal analysis of cytokine gene expression and parasite load in PBMC in Leishmania infantum experimentally infected dogs

Canine visceral leishmaniasis (CVL) is caused by Leishmania infantum, an intracellular protozoan parasite that causes a severe infectious disease. To evaluate the gene expression profile associated to CVL in vivo, we have measured monthly by real-time PCR over one year the IL-4, IL-10, IL-12, IL-13, IFN-gamma, TGF-beta and TNF-alpha mRNA levels in peripheral blood mononuclear cells in 6 experimentally infected dogs that exhibited different progressions of the illness. While in two dogs no parasite, or a very low number of parasites, was detected and the two dogs did not show any clinico-pathological abnormalities at the end of the study (L dogs), for the remaining dogs high parasite loads were detected and they developed clinical leishmaniasis (H dogs). The L dogs have null expression of both IL-4 and IL-13 for the first 4 months after the infection, whereas an early IL-4 and IL-13 expression occurs in this period of infection in most of the dogs that developed clinical leishmaniasis (H dogs). Furthermore, a higher IFN-gamma expression was associated with the increase of parasite load and clinical status in these dogs. Moreover, the high variability of expression at the pre-infection stage causes us to reject the possibility that the basal levels of these cytokines indicate the prognosis of the subsequent response against infection.     

Detection of Leishmania infantum by real-time PCR in a canine blood bank

O bjectives : Risk for transmission of Leishmania infantum from blood products has been largely demonstrated in human and veterinary literature. Appropriate screening of canine blood donors is important especially in an endemic area such as Barcelona (Spain). The purpose of this study was to evaluate the presence of L infantum DNA parasites by real-time quantitative PCR in our canine blood bank.
M ethods : Samples from blood products obtained from 92 canine blood donors were assayed for L infantum by means of real-time PCR amplification and quantification.
R esults : The prevalence of quantitative PCR-positive blood samples among healthy seronegative blood donors was 19·6 per cent.
C linical S ignificance : The results of this study show that L infantum infection is common in canine blood donors and their blood products in an endemic area, despite a negative commercial serological screening for infectious diseases. Therefore, screening by PCR should be included in an integrated approach to evaluate L infantum infection among potential blood donors.     

Pregnancy-related diabetes mellitus in two dogs

CASE SUMMARIES: Two cases of diabetes mellitus occurring in bitches in association with pregnancy are reported. In the first case, a bitch with suspected acromegaly developed diabetes mellitus within 2 weeks of the due date. Despite insulin therapy, euglycaemia was not achieved. Tw o live, small pups were delivered by elective Caesarean section but died within 2 days. Signs consistent with acromegaly resolved but diabetes mellitus was permanent in the bitch. In the second case, diabetic ketosis with severe gastrointestinal disease was diagnosed 2 days after Caesarean section was performed due to dystocia. The pups delivered all died within 5 days. The bitch recovered fully from diabetes mellitus within 2 weeks and has remained euglycaemic without insulin for a period of at least 18 months.

CLINICAL RELEVANCE: These two cases demonstrate that diabetes mellitus can occur in association with pregnancy in dogs, that diabetic ketosis can occur during transient diabetes mellitus in dogs, and suggest that acromegaly may occur during pregnancy-related dioestrus in dogs. The scarcity of previous reports of this nature, however, suggests that such cases are unusual.

Lack of prompt resolution of hyperglycaemia may result in secondary diabetes mellitus becoming permanent. Management should focus on immediate insulin therapy or ovariohysterectomy to minimise this risk. Even mild hyperglycaemia should not be ignored during pregnancy. The insulin antagonistic effects of pregnancy, stressful illness, surgery and dystocia can be enough to result in diabetic ketosis in the absence of permanent insulin deficiency. Maternal hyperglycaemia may contribute to adverse fetal outcomes in dogs but further study is required regarding the nature of the risk.     

Biology, pathogenicity, diagnosis and control of Ancylostoma caninum]

Infections with Ancylostoma caninum are transmitted orally or percutaneously. The transmission of infectious stages with the milk of particular importance for the distribution of the species. It occurs during the dissemination of larvae that follows every infection as well as after reactivation of resting somatic larvae in the bitch at the end of the pregnancy. The galactogenic transmission of larvae occurs even when, due to existing immunity, no patent infections develop in the bitch. Immunity does not or only to a low extent influence impatient infections or the migration of reactivated somatic larvae. It also allows a limited reestablishment of a deposit of larvae in the bitch. Following percutaneous infection dermatitis occurs in the area of larval penetration and the lung is affected by migrating larvae. Intestinal stages of Ancylostoma caninum damage the host by ingestion of the mucosa of the small intestine and withdrawal of blood. Main symptoms of ancylostomiasis are a mucous haemorrhagic diarrhoea and anaemia, that become visible 8 to 10 days post infection. The examination for impatient infections with Ancylostoma caninum can be done by immunofluorescence and ELISA. With both methods antibodies against third stage larvae can be detected from the first or second week post infection onward. Patent infections with Ancylostoma caninum can easily be detected by faecal examination for the presence of the characteristic oval, thin-walled eggs containing few blastomeres. Galactogenic infections with Ancylostoma caninum can be prevented or reduced by a regular treatment of the bitch with albendazole, fenbendazole or oxfendazole during the activation of larvae in the last third of the pregnancy or by repeated treatment with ivermectin shortly before and after birth. To prevent patent infections, galactogenic infected puppies have to be treated early and repeatedly.     

Pregnancy-related diabetes mellitus in two dogs

CASE SUMMARIES: Two cases of diabetes mellitus occurring in bitches in association with pregnancy are reported. In the first case, a bitch with suspected acromegaly developed diabetes mellitus within 2 weeks of the due date. Despite insulin therapy, euglycaemia was not achieved. Two live, small pups were delivered by elective Caesarean section but died within 2 days. Signs consistent with acromegaly resolved but diabetes mellitus was permanent in the bitch. In the second case, diabetic ketosis with severe gastrointestinal disease was diagnosed 2 days after Caesarean section was performed due to dystocia. The pups delivered all died within 5 days. The bitch recovered fully from diabetes mellitus within 2 weeks and has remained euglycaemic without insulin for a period of at least 18 months. CLINICAL RELEVANCE: These two cases demonstrate that diabetes mellitus can occur in association with pregnancy in dogs, that diabetic ketosis can occur during transient diabetes mellitus in dogs, and suggest that acromegaly may occur during pregnancy-related dioestrus in dogs. The scarcity of previous reports of this nature, however, suggests that such cases are unusual. Lack of prompt resolution of hyperglycaemia may result in secondary diabetes mellitus becoming permanent. Management should focus on immediate insulin therapy or ovariohysterectomy to minimise this risk. Even mild hyperglycaemia should not be ignored during pregnancy. The insulin antagonistic effects of pregnancy, stressful illness, surgery and dystocia can be enough to result in diabetic ketosis in the absence of permanent insulin deficiency. Maternal hyperglycaemia may contribute to adverse fetal outcomes in dogs but further study is required regarding the nature of the risk.     

The first record in the Americas of an autochthonous case of Leishmania (Leishmania) infantum chagasi in a domestic cat (Felix catus) from Cotia County, São Paulo State, Brazil

A case of feline cutaneous leishmaniasis is reported in a domestic cat (Felis catus) as an apparently natural infection in a non-endemic area. Amastigotes were seem in smears of a nodular lesion on the cat's nose. No parasite could be seen in cytological preparations of liver or spleen but DNA obtained from a sample of the spleen produced the expected fragment in a Leishmania specific rDNA based PCR assay. The PCR product, a 520 bp fragment, was sequenced and the nucleotide sequence was identical to that of Leishmania (Leishmania) infantum chagasi. These results are surprising since no autochthonous human or canine cases of visceral leishmaniasis have ever been reported from this region. This case suggests that natural transmission of this disease is occurring in this area, and that cats could act as a reservoir of L. (L.) infantum chagasi.     

Gestational diabetes mellitus with diabetic ketoacidosis in a Yorkshire terrier bitch

A 6 yr old pregnant Yorkshire terrier bitch presented 62 days after mating with an acute history of vomiting and coughing. The owners also reported that the dog was polyuric and polydypsic for the last 2 weeks. Complete blood count, serum biochemistry, and urinalysis revealed hyperglycemia, ketonemia, ketonuria, and metabolic acidosis. Diabetic ketoacidosis was diagnosed and after emergency treatment, including fluid therapy, prophylactic antibiotics, and regular insulin, the bitch whelped six healthy normal puppies. Two weeks after treatment, the bitch was clinically normal with normal fructosamine levels. To the authors' knowledge, this is the first reported case of gestational diabetes mellitus in a small breed dog.     

Little evidence of seasonal variation of natural infection by Leishmania infantum in dogs in Spain

Leishmania infantum, the etiological agent of canine leishmaniosis in the Mediterranean region, is vectored by Phlebotomus spp sandflies, which are active during the warmer months of the year. In order to determine whether seasonality in transmission induces seasonal changes in the prevalence of infection by L. infantum and of parasite-specific immune response, two groups of dogs, one in February (n=37) and another in October (n=42), were studied. Clinical signs compatible with leishmaniosis, as well as presence of microscopic skin lesions in the muzzle were recorded for all dogs. Assays were also performed for detection of L. infantum parasites in muzzle skin samples (PCR, immunohistochemistry and culture), specific serum antibodies (ELISA), and specific lymphocyte proliferation and interferon-gamma production. Although prevalence of non-specific clinical signs increased significantly after the sandfly season, this was not the case for Leishmania-specific markers: positivity by PCR (24% vs. 21%) or immunohistochemistry (3% vs. 2%) of muzzle skin samples, as well as lymphocyte proliferation (59% vs. 50%) or interferon-gamma production (21% vs. 27%) were similar in February and in October. Only prevalence of positive specific antibody titers increased noticeably in October (8% vs. 20%), although this was not statistically significant. Overall, the sandfly season did not have a marked impact on the prevalence L. infantum infection or parasite-specific immune responses analyzed in this study.     

Urological complications following ovariohysterectomy in a bitch

A two-year-old chihuahua bitch was presented with urinary incontinence following ovariohysterectomy which had been performed at the time of caesarean section. Clinical, contrast radiographic and surgical investigation revealed the presence of right hydroureter and hydronephrosis, vesicovaginal fistula and absence of most of the urinary bladder. These abnormalities appeared to have resulted from ligation of the caudal bladder, at the level of the trigone, during ovariohysterectomy.     

Phosphorus deficiency in cattle on two farms in canterbury

A New Zealand canine herpesvirus isolate was inoculated into three 2-day-old puppies via the intraperitoneal route, two other puppies from the same litter being retained as in-contact controls. All pups were left suckling the bitch.

One inoculated pup died of misadventure. The remaining inoculated pups, and one in-contact pup, died with clinical signs of herpesvirus infection, the virus being subsequently recovered from a number of tissues. The remaining in-contact pup also developed typical disease, but recovered, virus being detected only in the tonsils. Lesions were detected in the diseased puppies in a wide variety of organs, and were consistent with previously published reports. No evidence of disease was detected in the bitch, but both she and the recovered pup developed neutralizing antibodies to the virus.     

Estimated pregnancy length from ovulation to parturition in the bitch and its influencing factors: a retrospective study in 162 pregnancies

An accurate timing of parturition is very useful for managing canine parturition. It is generally accepted that parturition in bitches occurs between 64 and 66 days after the luteinizing hormone peak. In this retrospective study, we determined pregnancy length in different breeds and its influencing factors dating it from the estimated day of ovulation (EDO), defined as the day when peripheral plasma level of progesterone (P4) reaches 6 ng/ml. From January 2001 to December 2006, 162 pregnancies in 151 bitches of 53 different breeds were followed. Different parameters concerning the bitch, the litter, the type of semen and the type of reproduction were studied. The mean estimated pregnancy length in the bitch from EDO to parturition was 63.1±2.1 days. The main influencing factors for the pregnancies studied were the breed, the size of the bitch and the number of puppies within the litter.     

Prolonged foetal retention in a bitch resulting in trichogranulomatuous panmetritis and re-establishment of fertility after unilateral ovariohysterectomy.

This case report describes the clinical course, the diagnostic and therapeutic management as well as histopathological findings in a case of foetal retention for about 5 months in a 5-year-old Boxer. Clinical symptoms included mucosanguinous vaginal discharge and a firm ovoid mass of about duck egg size palpable in mid abdomen. Sonographic examination was not suitable to visualize uterine contents, while radiography revealed a conglomeration of foetal bones focally visible in the right uterine horn, which was surgically removed together with the ipsilateral ovary in order to maintain the fertility of the bitch. After lancing the uterine wall the osseous skeleton as well as parts of hairy skin and muscles of the foetus attaching the uterine wall were detected. Histologically a unilateral necrotizing panmetritis with almost complete destruction of the endometrial structures was diagnosed. Furthermore, the endometrium, the myometrium as well as the serosa revealed a severe trichogranulomatuous inflammation with numerous foetal hairs visible within the inflammatory alterations. After treatment of the bitch with prostaglandin F(2alpha) and an antibiotic agent she became pregnant in the second heat after surgery and gave birth to two healthy puppies. This is the first report on foetal retention for several months followed by unilateral ovariohysterectomy with subsequent recovery of fertility. Furthermore, this case report describes for the first time the histomorphological diagnosis of a unilateral trichogranulomatuous panmetritis in a bitch.     

Evaluation of a new non-hormonal substance for terminating pregnancy in bitches in clinical trials

A new non-hormonal contragestational agent (DL 717-IT), the 2-(4 chlorophenyl)-s-triazole |5,1-a|-isoquinoline, was tested in clinical trials to determine its efficacy for interrupting the pregnancy in the bitch. Five hundred and sixty-five bitches were given a single intramuscular injection of the drug between the first and 15th day after the mating. Of the 475 bitches that received 2–5 mg/kg, 430 (90-5 per cent) did not whelp at all, 15 (3-2 per cent) delivered a few stillborn or weak puppies that died some hours after birth, and 30 (6-3 per cent) delivered at least one normal live pup. Of 90 bitches treated with 3–5 mg/kg, 87 (96-7 per cent) did not whelp at all, two (2-2 per cent) delivered a few stillborn or weak puppies that died some hours after birth, and one (1 1 per cent) delivered normal live puppies. DL 717-IT, owing to the high effectiveness and minimal side effects, must be considered a very reliable drug for the interruption of pregnancies in the bitch.