Administration of aromatase inhibitor MPV-2213ad to blue fox vixens (Vulpes lagopus) as a model for contraception in female dogs.

The interest in non-surgical approaches to contraception and fertility control in female dogs has increased in recent years. In this study the effect of an aromatase inhibitor (finrozole) was evaluated in fur production animals, farmed blue fox vixens, as a model for contraception in bitches. A total of 80 vixens were divided into 4 groups, receiving orally placebo (A) or finrozole 0.5 mg/kg (B), 3.5 mg/kg (C) or 24.5 mg/kg (D) for 21 consecutive days beginning in the pre-ovulatory period of heat. Monitoring of the vixens included clinical signs of heat, measurement of vaginal electrical resistance (VER) as well as oestradiol and progesterone concentrations in plasma. The approximate relation of the start of treatment to ovulation varied from 11 days before to one day after ovulation provided that the LH peak occurred 0.5-2 days before the VER peak and ovulation was then estimated to occur 2 days after the LH peak. Seventy vixens were artificially inseminated within 8 h after a 50 Ω decline in vaginal electrical resistance was detected. Ten vixens were not inseminated. Pregnancy was confirmed by transabdominal ultrasound examination and birth of cubs was recorded. The pregnancy rates in the groups were 89.5% (A), 81.3% (B), 55.6% (C) and 52.9% (D). The average number of live born pups in the four groups was 9.4 (A), 7.0 (B), 5.8 (C), and 3.8 (D), respectively. No deleterious effects (for instance malformations) of finrozole on pups could be verified. The administration of finrozole did not have a significant effect on oestradiol parameters and VER values in vixens. Progesterone values were significantly higher in treatment groups compared with the placebo group. The results indicate that pregnancy could be avoided by finrozole provided that doses of ≥3.5 mg/kg were used and the treatment was initiated at least four days before the day of artificial insemination. This corresponds with two to six days before ovulation provided that the LH peak occurred 0.5-2 days before the VER peak and that ovulation then occurred in average 2 days after the LH peak.

Polyorchidism with presumed contralateral intrauterine testicular torsion.

INTRODUCTION: Polyorchidism was first described by Blasius in 1670(1) during a routine autopsy. We report a child with unilateral polyorchidism and a contralateral absent testis, a combination not reported previously. PRESENTATION OF CASE: A 2-year-old boy was referred to the outpatient clinic with an impalpable left testis. At laparoscopy, the left vas deferens and testicular vessels ended blindly proximal to a closed internal ring. No gonadal tissue was identified. On the right side, a single vas deferens and testicular vessels were seen entering the internal ring as normal. The right side of the scrotum was explored and two testes were identified within a single tunica vaginalis. DISCUSSION: Polyorchidism is rare with a literature search identifying approximately 230 reported cases. Whilst prenatal testicular torsion is increasing being recognized and treated as a surgical emergency,(9) prenatal testicular torsion in association with polyorchidism has not been previously reported. CONCLUSION: We describe a unique case of a 2-year-old boy with right-sided polyorchidism and an absent left testis associated with a blind ending vas deferens and testicular vessels, presumed secondary to intrauterine testicular torsion.

Pathogen exclusion properties of canine probiotics are influenced by the growth media and physical treatments simulating industrial processes.

AIMS: Manufacturing process used in preparation of probiotic products may alter beneficial properties of probiotics. The effect of different growth media and inactivation methods on the protective properties of canine-originated probiotic bacteria against adhesion of canine enteropathogens was investigated. METHODS AND RESULTS: Three established dog probiotics, Lactobacillus fermentum VET9A, Lactobacillus plantarum VET14A and Lactobacillus rhamnosus VET16A, and their mixture were assessed using the dog mucus pathogen exclusion model. The pathogens used were Enterococcus canis, Salmonella enterica serovar Typhimurium and Clostridium perfringens. The effect of growth media, one reflecting laboratory and the other manufacture conditions, and viability (viable and heat inactivated, 80°C per 30 min) on the pathogen exclusion properties of probiotics were characterized. Greater pathogen exclusion percentages were noted for probiotics growing in conditions reflecting manufacture when compared to laboratory (P < 0·05). Inactivation of probiotics by heat (80°C per 30 min) increased pathogen exclusion compared with their viable forms (P < 0·05). CONCLUSIONS: Manufacturing process conditions such as growth media, incubation temperature and pretreatment methods may significantly affect the protective properties of the tested strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Growing conditions and pretreatment methods should be carefully considered when designing new probiotics to reduce the risk of common infections in dogs. The studied probiotics are promising potential feed additives for dogs.

The effect of growth media and physical treatments on the adhesion properties of canine probiotics.

AIMS: The manufacturing processes have been reported to influence the properties of probiotics with potential impact on health properties. The aim was to investigate the effect of different growth media and inactivation methods on the properties of canine-originated probiotic bacteria alone and in combination mixture. METHODS AND RESULTS: Three established dog probiotics, Lactobacillus fermentum VET9A, Lactobacillus plantarum VET14A and Lactobacillus rhamnosus VET16A, and their combination mixture were evaluated for their adhesion to dog mucus. The effect of different growth media, one reflecting laboratory and the other manufacturing conditions, and inactivation methods (95°C, 80°C and UV irradiation) on the mucus adhesion of the probiotic strains was characterized. Evaluation of dog probiotics was supported by cell visualization using transmission electron microscopy (TEM). Higher adhesion percentage was reported for probiotic strains growing in laboratory rather than in manufacturing conditions (P < 0.05). Inactivation by heat (95°C, 80°C) decreased the adhesion properties when strains were cultivated in soy-based growth media compared with those grown in MRS broth (P < 0.05). TEM observations uncovered differences in cell-surface components in nonviable forms of probiotic strains as compared with their viable forms. CONCLUSIONS: Manufacturing process conditions such as growth media and pretreatment methods may significantly affect the adhesive ability of the tested strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Growth conditions, growth media, pretreatment methods and different probiotic combinations should be carefully considered for quality control of existing probiotics and for identification of new probiotics for dogs. These may also have an impact on health benefits for the host.

Excitatory amino acid transporter 5 is widely expressed in peripheral tissues.

It is routinely stated in the literature that Excitatory Amino Acid Transporter 5 (EAAT5) is a retina-specific glutamate transporter. EAAT5 is expressed by retinal photoreceptors and bipolar cells, where it serves as a slow transporter and as an inhibitory glutamate receptor, the latter role is due to the gating of a large chloride conductance. The dogma of an exclusively retinal distribution has arisen because Northern blot analyses have previously shown only modest hybridisation in non-retinal tissues. Others have re-interpreted this as indicating that EAAT5 was only present in retinal tissues. However, this view appears to be erroneous; recent evidence demonstrating abundant expression of EAAT5 in rat testis prompted us to re-examine this dogma. A new antibody was developed to an intracellular loop region of rat EAAT5. This new tool, in concert with RT-PCR and sequencing, demonstrated that EAAT5 is widely distributed at the mRNA and protein levels in many non-nervous tissues including liver, kidney, intestine, heart, lung, and skeletal muscle. We conclude that EAAT5 is a widely distributed protein. Whether it functions in all locations as a glutamate transporter, or mainly as a glutamate-gated chloride conductance, remains to be determined.

Establishing benchmarks for the outcome of herniotomy in children.

BACKGROUND: Although complication rates following herniotomy in children are low, reported rates vary. The aim of this study was to propose desirable outcome measures that can be used by surgeons to assess the quality of their service. METHODS: A review of prospectively collected data on all hernias operated on by the Christchurch-based paediatric surgical service was undertaken. Complication rates were determined and compared with those reported in the literature. Rates for children aged 1 year or above were compared with those in children aged less than 1 year. RESULTS: There were 3128 herniotomies performed, with an overall recurrence rate of 0.6 per cent. Wound infections occurred in 0.9 per cent, but were confined to infants aged less than 12 months. The rate of acquired cryptorchidism was 0.4 per cent and testicular atrophy 0.8 per cent in boys. Injury to the vas deferens was recognized in 0.1 per cent. Children under 1 year of age were more likely to develop recurrence than older children (0.9 versus 0.2 per cent respectively; odds ratio 4.56, P = 0.013). CONCLUSION: Complication rates were relatively low compared with those reported previously, but were considered achievable in most paediatric surgical units. Specific attention should be paid to the youngest infants, as this group has the highest complication rate. Minimum standards can be proposed based on these and other published outcomes.

The degree to which the size and side of an inguinal hernia is predictive of a hernia on the other side.

BACKGROUND: The indication for contralateral exploration in patients with a unilateral inguinal hernia and the implications of a contralateral patent processus vaginalis have been the subject of much debate during recent decades. The aim of this study was to investigate whether the size and side of the hernial sac on the side of presentation are predictive for the operative appearance of the processus vaginalis/hernial sac on the contralateral side. METHODS: All primary herniotomies performed by the Christchurch-based paediatric surgical service were reviewed. Patients who developed a contralateral hernia after unilateral repair were identified. In a random subset of patients who had bilateral exploration for unilateral hernias, the size of the clinical hernial sac was plotted against the size of the contralateral patent processus vaginalis/hernial sac (when present) as assessed at the time of open surgery. RESULTS: Overall, 2,124 unique patients underwent herniotomies in the period reviewed, and 44 (3.8%) metachronous contralateral hernias were identified. Girls were not at greater risk than boys of developing a metachronous hernia. There was a positive correlation between the size of the clinical hernial sac and the size of the contralateral processus vaginalis/hernial sac. There was no significant relationship between the original presenting side and the development of a metachronous contralateral hernia. CONCLUSION: It is not possible to predict reliably which patients who present with a unilateral hernia would benefit from bilateral exploration, although the size of the hernia has a positive correlation with the size of the contralateral processus vaginalis/hernial sac. Those with "massive" hernias on the presenting side were more likely to have "large" contralateral hernial sacs, but how many of these would subsequently become symptomatic remains uncertain.

Lactic acid bacteria isolated from canine faeces.

AIMS: Lactic acid bacteria (LAB) were isolated and sequenced from the faeces of healthy dogs. Five of these strains were selected and further characterized to clarify the potential of these strains as probiotics for canine. METHODS AND RESULTS: LAB were found in 67% (14/21) of the canine faeces samples when plated on Lactobacilli Selective Media without acetic acid. Out of 13 species identified with partial 16S rRNA gene sequencing, Lactobacillus fermentum LAB8, L. mucosae LAB12, L. rhamnosus LAB11, L. salivarius LAB9 and Weissella confusa LAB10 were selected as candidate probiotic strains based on their frequency, quantity in faeces, growth density, acid tolerance and antimicrobial activity. The minimal inhibitory concentration values of these isolates were determined for 14 antibiotics. L. salivarius LAB9, W. confusa LAB10 and L. mucosae LAB12 were viable in pH 2 for 4 h (mLBS), indicating tolerance to acidity and thus the potential to survive in gastrointestinal tract of the canine. The LAB8-LAB12 strains showed antimicrobial activity against Micrococcus luteus A1 NCIMB86166. CONCLUSIONS: Thirteen different LAB species were found from the faecal microbiota of the healthy canines. Five acid tolerant and antimicrobially active LAB strains with the capacity to grow to high densities both aerobically and anaerobically were chosen to serve as candidate probiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: The selected LAB strains are among the first host-specific LAB with antimicrobial activity isolated from canines that could serve as potential probiotics for canine use.

Intussusception and rotavirus associated hospitalisation in New Zealand.

AIMS: To describe the epidemiology of intussusception and its relation to rotavirus associated hospitalisation in New Zealand. METHODS: National hospital discharge data between January 1998 and June 2003 for all children younger than 3 years of age with intussusception were reviewed. Independently, children from the same age group, admitted to eight paediatric units with rotavirus gastroenteritis between May 1998 and May 2000, were identified prospectively. Epidemiological characteristics of cases with intussusception were compared with those of hospitalised rotavirus disease. RESULTS: During the 5.5 year study period, there were 277 cases of intussusception and no deaths. Most (72%) occurred in the first year of life (age adjusted incident rate 65 per 100,000 child-years, 95% CI 56 to 74). Risk of intussusception was less in females (risk ratio 0.58; 95% CI 0.43 to 0.78) and for Maori (risk ratio 0.52; 95% CI 0.35 to 0.77) when compared with European infants. In contrast to hospitalised rotavirus cases, intussusception peaked at a younger age and lacked seasonality. CONCLUSIONS: This study provides national baseline data on intussusception for future rotavirus vaccine programmes in New Zealand. Wild-type rotaviruses do not appear to have a major role in triggering intussusception. Prospective surveillance systems, using standardised case definitions and nested case-control methodology, are needed to further our understanding of the aetiology and epidemiology of intussusception.

The development of the proximal oesophageal pouch in the adriamycin rat model of oesophageal atresia with tracheo-oesophageal fistula.

This study examined the morphological development of the proximal oesophagus in the Adriamycin-induced rat model of oesophageal atresia. The proximal oesophageal segment in oesophageal atresia with tracheo-oesophageal fistula (OA\TOF) has been assumed to be of similar embryological origin to the distal oesophagus. However, recent research using the Adriamycin model of OA\TOF has indicated that these structures may have a different origin. Time-mated Sprague-Dawley rats were administered either Adriamycin intraperitoneally or saline of an equivalent volume between days 6-9 of gestation. The rats were sacrificed between days 11-19 of gestation, their embryos removed and histologically sectioned. These sections were analysed to observe the morphological changes occurring in the proximal foregut. The proximal oesophageal pouch first appeared on day 15.25 as a dorsal outpouching of the proximal foregut immediately cranial to an area of apoptosis in the dorsal epithelium of the distal pharynx. It elongated through a process of cellular proliferation until it was clearly formed on day 16. Relatively little growth occurred from days 17-19. In the rat developing oesophageal atresia, the proximal oesophageal pouch has an origin different to that of the distal oesophagus. This study may explain the difference in immunohistological properties and intrinsic nervous supply between the proximal and distal oesophageal segments in oesophageal atresia.

Characterization and electrotransformation of Lactobacillus crispatus isolated from chicken crop and intestine.

Lactic acid bacteria originating in the intestine have recently undergone intensive study for their potential probiotic properties. Here partial 16S rRNA gene sequencing of 8 Lactobacillus strains proved them to be Lactobacillus crispatus. Fatty acid analysis confirmed strains being closely related. These strains and type strain ATCC33820 were characterized for genetic engineering potential, thus determining aerobic growth, erythromycin sensitivity, and glycine tolerance. Out of 5 plasmids, a 2.9-kb plasmid (pLEB579) was successfully introduced into 4 chicken-originated wild-type L. crispatus strains. Transformation frequency was approximately 30 transformants per microgram of DNA, the first reported electrotransformation into chicken-originated L. crispatus. In spite of its low frequency, transformation enables bioengineering of these strains to improve the probiotic function in feed adsorption, chicken health, and food safety.

Fatal late onset necrotising enterocolitis in a term infant: Atypical Kawasaki disease or polyarteritis nodosa of infancy?

A 3-month-old infant died following a 3-week illness that commenced with diarrhoea and vomiting, and produced progressive infarction of his small bowel. Post-mortem identified multiorgan arteritis of uncertain aetiology, giving rise to coronary artery occlusion and myocardial infarction as well as necrosis of the entire small bowel and other organs. Kawasaki disease and polyarteritis nodosa of infancy are proposed as possible aetiologies in this case.

Fermented soymilk with a monoculture of Lactococcus lactis.

Lactococcus lactis strain (LL3) isolated from mothers' milk was used to produce fermented soymilk. The strain survived at levels of over 7 log cfu/ml for 3 weeks in the fermented soymilk. A consumer survey was carried out to compare the acceptability of the fermented product with a similar product made with L. lactis ATCC11545 originally isolated from cow's milk. Blind samples produced by fermentation with the two strains were rated equally attractive, whereas information on the origin of the strains significantly enhanced the pleasantness of the fermented soymilk.

Familial parathyroid carcinoma in a child.

Familial hyperparathyroidism and parathyroid carcinoma are rare diseases. A case of parathyroid carcinoma in an 8-year-old girl whose mother had previously undergone parathyroidectomy for primary hyperparathyroidism is reported. Parathyroid carcinoma in a preadolescent child has not been described previously, and may have a familial basis.

Abnormal branching and regression of the notochord and its relationship to foregut abnormalities.

An abnormally positioned notochord has been reported in embryos that develop foregut abnormalities, vertebral defects and other abnormalities of the VATER association. This study examines the patterns of regression of the abnormal notochord in the rat model of the VATER association and investigates the relationship between developmental abnormalities of the notochord and those of the vertebra and foregut. Timed-pregnant Sprague-Dawley rats were given daily intraperitoneal injections of 1.75 mg/kg adriamycin on gestational days 6 - 9 inclusive. Rats were sacrificed between days 14 and 20 and their embryos harvested, histologically sectioned and stained and examined serially. The location and appearance of the degenerating notochord and its relationship to regional structural defects were analysed. All 26 embryos exposed to adriamycin developed foregut abnormalities and had an abnormal notochord. The notochord disappeared by a process of apoptotic degeneration that lagged behind that of the normal embryo: the notochord persisted in the abnormal embryo beyond day 17, whereas in the normal rat it had already disappeared. Similarly, formation of the nucleus pulposus was delayed. Vertebral abnormalities occurred when the notochord was ventrally-positioned. The notochord disappears during day 16 in the normal embryo whereas abnormal branches of the notochord persist until day 19 in the adriamycin-treated embryo. Degeneration of the notochord is dominated by apoptosis. An excessively ventrally-placed notochord is closely associated with abnormalities of the vertebral column, especially hemivertebrae.

Hedgehog in the human: a possible explanation for the VATER association.

Foregut malformations are relatively common anomalies, occurring in 1 in 2000-5000 live births. The adriamycin-induced rat model of the VATER association has provided a means of studying the morphogenesis of a variety of major congenital structural abnormalities similar to those seen in humans with VATER association. The secreted glycoprotein, Sonic hedgehog (Shh), may act as an endodermal signal that controls gut and lung patterning. Mice with targeted deletion of Shh have foregut defects that are consistent with those produced by administration of adriamycin. It is possible that mutations induced by adriamycin may result from the breakdown of the Shh signalling pathway.