The transmission potential of Rift Valley fever virus among livestock in the Netherlands: a modelling study

Abstracts

Rift Valley fever virus (RVFV) is a zoonotic vector-borne infection and causes a potentially severe disease. Many mammals are susceptible to infection including important livestock species. Although currently confined to Africa and the near-East, this disease causes concern in countries in temperate climates where both hosts and potential vectors are present, such as the Netherlands. Currently, an assessment of the probability of an outbreak occurring in this country is missing. To evaluate the transmission potential of RVFV, a mathematical model was developed and used to determine the initial growth and the Floquet ratio, which are indicators of the probability of an outbreak and of persistence in a periodic changing environment caused by seasonality. We show that several areas of the Netherlands have a high transmission potential and risk of persistence of the infection. Counter-intuitively, these are the sparsely populated livestock areas, due to the high vector-host ratios in these areas. Culex pipiens s.l. is found to be the main driver of the spread and persistence, because it is by far the most abundant mosquito. Our investigation underscores the importance to determine the vector competence of this mosquito species for RVFV and its host preference.     

Dominant expression of interleukin-8 vs interleukin-1β and tumour necrosis factor alpha in lungs of lambs experimentally infected with Mannheimia haemolytica

Abstract

AIMS: To quantify the number of cells infected with Mannheimia haemolytica and expressing interleukin (IL)-1β, tumour necrosis factor alpha (TNFα) and IL-8 using immunohistochemistry, and to measure the immunoreactivity of cytokines in pulmonary tissue extracts using ELISA, in the lung of lambs experimentally infected with M. haemolytica, and to compare the patterns of expression of cytokines in airways at different times post-infection (p.i.).

METHODS: Twenty 3-month-old lambs of both sexes were randomly assigned to two groups, viz infected (n=15), and uninfected controls (n=5). Each lamb in the infected group was inoculated with 1.5 x 10⁹ cfu M. haemolytica in 5 mL sterile nutrient broth, control lambs were inoculated with 5 mL sterile nutrient broth and clinical signs were monitored. Infected and control animals were killed at 1, 3, 5, 7, and 15 days p.i. Histopathology and immunohistochemistry were conducted to determine the number of immunolabelled cells in pneumonic lungs, and study the pattern of expression of IL-1β, TNFα and IL-8 in lung extracts using ELISA.

RESULTS: Lesions in bronchi and bronchioles ranged from epithelial desquamation to bronchiolitis obliterans and necrosis. The alveoli had areas of seroproteinaceous fluid, fibrin and bacterial aggregates that evolved to foci of pyogranulomatous inflammation with clustered inflammatory cells, referred to as ‘oat cells’. M. haemolytica antigen was observed in the cytoplasm of inflammatory cells. Labelling of IL-1β, TNFα and IL-8 was observed in bronchial and bronchiolar epithelial cells, alveolar exudate, and in interstitial inflammatory infiltrate, with increased expression on 1 and 3 days p.i. for IL-1β and TNFα, and 1, 3, and 5 days p.i. for IL-8. In lung tissue extracts, peak concentrations of IL-1β (55 (SD 5) ng/mL), TNFα (92 (SD 6) pg/mL) and IL-8 (8 [SD 2] μg/mL) occurred at 3 days p.i.

CONCLUSIONS: The results of this study suggested that the inflammatory cytokines IL-1β, TNFα and IL-8 may play an important role in enhancing the biological response to M. haemolytica, and contribute to the development of lesions in the lung in pulmonary pasteurellosis in sheep. Given that the expression of IL-8 in lung was much greater than that of IL-1β and TNFα, anti-cytokine agents directed at this mediator could be useful in the prevention and treatment of this disease.     

Effect of hCG and eCG Treatments on Prostaglandins Synthesis in the Porcine Oviduct

The oviduct plays a crucial role in fertilization, gamete maturation and embryo transport. Prostaglandins are some of the main factors determining its roles. The present study investigated the influence of oestrus synchronization and superovulation on prostaglandins synthesis in the porcine oviduct. Mature cross‐bred gilts after exhibiting oestrous cycles were divided into four groups: I, cyclic; II, inseminated; III, synchronized and inseminated; and IV, superovulated and inseminated. Oviducts were collected on the third day of the oestrous cycle or after insemination and divided into isthmus and ampullary parts. This study demonstrated lower mRNA (in the isthmus and ampulla; p < 0.05, p < 0.001, respectively) and protein prostaglandin endoperoxide synthase 2 expression (in the isthmus; p < 0.001) in gilts treated with human chorionic gonadotrophin/equine chorionic gonadotrophin (hCG/eCG) compared with Group II that were inseminated only. In addition, hCG and eCG treatment decreased mPGES‐1 mRNA levels in Groups III and IV, in both the isthmus (p < 0.01 in III, p < 0.001 in IV) and ampulla (p < 0.001). The prostaglandin E₂ content of oviductal tissues was significantly lower in Groups III (p < 0.05) and IV (p < 0.01 in isthmus, p < 0.0001 in ampulla) compared with Group II. mRNA and protein levels of PGFS in Group IV in the oviductal isthmus were higher (p < 0.01) compared with the non‐treated Group II. In effect, the amount of prostaglandin F_2α in oviductal tissues of gilts treated with hCG/eCG was significantly elevated (p < 0.001 in isthmus of Groups III and IV; p < 0.05 in ampulla of Group IV). Differential expression of the factors analysed in gilts treated with exogenous gonadotrophins suggests that hCG/eCG stimulation affects prostaglandins synthesis pathway. These changes can alter oviduct functions and in turn affect gamete maturation and fertilization as well as development of embryos and their transport to the uterus.     

Effects of Vasectomy on Seminal Plasma Alkaline Phosphatase in Male Alpacas (Vicugña pacos)

Azoospermia is a common finding in male alpacas which present for infertility. The challenge is to differentiate azoospermia of testicular origin from non‐testicular origin. In several species, alkaline phosphatase (AP) concentrations in seminal plasma have been used as a diagnostic marker of contributions of the testis and epididymis to the ejaculate. The purpose of this study was to determine whether AP assay could differentiate testicular from non‐testicular azoospermia in male alpacas. An experimental model of bilateral outflow obstruction (pre‐scrotal vasectomy) was used in 22 male alpacas, aged 2–9 years. No reproductive history was available. Animals were submitted for electroejaculation (EE) under general anaesthesia and vasectomy performed. Five weeks later, animals were submitted for EE. Vasectomy was not successful in one animal, which was removed from analysis. AP levels were compared in seminal plasma in the pre‐ and post‐vasectomy samples. The mean ± SEM concentration of AP in pre‐vasectomy seminal plasma was 504.29 ± 166.45 U/l (range 10–2910); the post‐vasectomy levels were 252.48 ± 81.77 U/l (range 0–1640; p = 0.06). In 71.4% of animals, AP levels decreased, varying from 18% to 100% reduction. Results of this study suggest that AP is not produced exclusively by the testis and epididymis in alpacas and that AP assay is not a valid diagnostic test for determination of origin of azoospermia; the gold standard for diagnosis of origin of azoospermia remains testicular biopsy.     

Isolation and genotyping of Campylobacter species from kiwi (Apteryx spp.) in captivity: implications for transmission to and from humans

Aims: To investigate the presence of Campylobacter spp. in captive kiwi (Apteryx spp.) and compare their genotypic profiles with those of human and animal origin, in order to assess their potential for zoonotic or zooanthroponotic transmission.

Methods: Conventional selective enrichment and filter-based isolation methods were applied to isolate Campylobacter spp. from fresh faecal samples from 12 North Island brown kiwi (Apteryx mantelli) and one great spotted kiwi (A. haastii), housed in one of five different areas in a kiwi sanctuary in Christchurch, New Zealand. Isolates were identified using multiplex PCR and 16S rRNA gene sequencing. High-resolution rapid genotyping using multiplex ligation-dependant probe amplification-based binary typing (MBiT) was applied and profiles compared with similar results from 2,165 Campylobacter spp. isolates contained in a database derived from human clinical, veterinary and environmental samples.

Results: One isolate of C. jejuni, and one belonging to the C. lari phylogenetic group were recovered from faeces from two kiwi. High-resolution rapid genotyping by MBiT demonstrated these to be indistinguishable from isolates obtained previously from human cases of diarrhoea, and others from chicken, cattle, sheep and water.

Conclusions: These data provide evidence for potential zoonotic or zooanthroponotic transmission of Campylobacter spp. in kiwi with implications for management of birds kept in captivity. We believe this is the first formal report of C. jejuni and a C. lari-like organism in kiwi.

Abbreviations: MBiT: Multiplex ligation-dependant probe amplification-based binary typing     

Significant association of serum autoantibodies to TYMS,HAPLN1 and IGFBP5 with early stage canine malignant mammary tumours

Canine mammary tumours (CMTs) are the most prevalent neoplasms in female dogs. Despite the high incidence of such tumours, a lack of easily accessible biomarkers still impedes early diagnosis of malignant CMTs. Herein we identify thymidylate synthetase (TYMS), hyaluronan and proteoglycan link protein 1 (HAPLN1) and insulin‐like growth factor‐binding protein 5 (IGFBP5) as CMT antigens eliciting corresponding autoantibodies in CMT cases. We establish enzyme‐linked immunosorbent assays (ELISAs) to detect autoantibodies to TYMS (TYMS‐AAb), HAPLN1 (HAPLN1‐AAb) and IGFBP5 (IGFBP5‐AAb) in sera from 81 dogs with malignant CMTs (41 in Stage I), 24 with benign CMTs and 35 healthy controls. Levels of all the three autoantibodies are elevated in the malignant group compared with the healthy or the benign group; notably, the elevated autoantibody levels significantly correlate with the stage‐I CMTs. For discriminating malignant CMTs from healthy control, the area under curve (AUC) of TYMS‐AAb is 0.694 with specificity of 82.9% and sensitivity of 50.6%. The AUC of utilising HAPLN1‐AAb for distinguishing the stage‐I CMTs from healthy controls is 0.711 with specificity of 77.1% and sensitivity of 58.5%. In differentiating malignant CMTs from the benign, the AUC of IGFBP5‐AAb reaches 0.696 with specificity of 70.8% and sensitivity of 67.9%, and a combination of IGFBP5‐AAb and TYMS‐AAb increases the AUC to 0.72. Finally, the AUC of combined HAPLN1‐AAb and IGFBP5‐AAb in discriminating the stage‐I CMTs from the benign achieves 0.731. Collectively, this study highlights a significant association of the three serum autoantibodies with early stage malignant CMTs.     

Persistent isolated hypocortisolism following brief treatment with trilostane

A 12‐year‐old male neutered Miniature Poodle with confirmed pituitary‐dependent hyperadrenocorticism was treated with trilostane. After three doses, it developed clinical and laboratory changes suggestive of isolated hypocortisolism (‘atypical hypoadrenocorticism’), which persisted and progressed for more than 3 months despite immediate withdrawal of the trilostane. The clinical signs of hyperadrenocorticism resolved without further trilostane. After 3 months, prednisolone treatment was started and the clinical signs of hypocortisolism resolved. Prednisolone therapy was required for more than 1 year. Ultrasonography initially demonstrated large hypoechoic adrenal cortices, typical of dogs with hyperadrenocorticism, which then became small and heteroechoic, consistent with the development of adrenal necrosis. Persistent isolated hypocortisolism has not been reported previously as a complication of trilostane therapy. The case is also remarkable for the very short duration of trilostane therapy that elicited this complication. Clinicians should be aware that trilostane therapy may result in adrenal necrosis, even in the very earliest stages of therapy, but prompt action can prevent a life‐threatening situation.     

Prospective evaluation of pancarpal arthrodesis for carpal injuries in working dogs in New Zealand,using dorsal hybrid plating

Abstract

AIM: To determine whether working dogs in New Zealand with carpal injuries and treated with unilateral pancarpal arthrodesis (PCA), using a dorsal hybrid-plating method, are able to return to satisfactory working ability.

METHODS: Fourteen working dogs presented to the Veterinary Specialist Group (VSG) and the Massey University Veterinary Teaching Hospital (MUVTH) with carpal injuries were prospectively treated using dorsal hybrid plating. Dogs were eligible if actively involved in farm, hunting or police work. Dogs had a standardised PCA surgical procedure performed, and similar instructions for post-operative care were provided. Dogs were re-evaluated clinically and radiographically at 6 weeks, 6 months, and 12 months after surgery. A questionnaire was completed by 12 owners, to assess each dog's working ability.

RESULTS: Twelve months following arthrodesis, 10/12 (83%) dogs could perform most or all duties normally. Eleven owners (92%) reported that the result of the surgery met their expectations, and nine owners (75%) were very satisfied with the outcome of the surgery. No owners were disappointed or very disappointed with the surgical outcome. Post-operative complications requiring surgical removal of the implant occurred in three (25%) dogs.

CONCLUSIONS: Unilateral PCA using a standardised surgical procedure and dorsal hybrid plating of carpal injuries has a good prognosis for working dogs in New Zealand to return to work.

CLINICAL RELEVANCE: These results may allow veterinarians to provide a more accurate prognosis to owners of working dogs that have debilitating carpal injury