Evaluation of post-exposure prophylaxis practices to improve the cost-effectiveness of rabies control in human cases potentially exposed to rabies in southern Bhutan.

BACKGROUND: Rabies is endemic in southern Bhutan, associated with 1-2 human deaths and high post exposure prophylaxis (PEP) costs annually. Evaluation of clinicians' management of human cases potentially exposed to rabies could contribute to improving PEP prescribing practices to both reduce unnecessary costs associated with PEP and reach the target of zero human deaths due to rabies by 2023. METHODS: A cross-sectional survey of 50 clinicians' management of human cases potentially exposed to rabies was conducted in 13 health centers in high-rabies-risk areas of Bhutan during February-March 2016. RESULTS: Data were collected on clinicians' management of 273 human cases potentially exposed to rabies. The 50 clinicians comprised health assistants or clinical officers (55%) and medical doctors (45%) with a respective median of 19, 21 and 2 years' experience. There was poor agreement between clinicians' rabies risk assessment compared with an independent assessment for each case based on criteria in the National Rabies Management Guidelines (NRMG). Of the 194 cases for which clinicians recorded a rabies risk category, only 53% were correctly classified when compared with the NRMG. Clinicians were more likely to underestimate the risk of exposure to rabies and appeared to prescribe PEP independently of their risk classification.. Male health assistants performed the most accurate risk assessments while female health assistants performed the least accurate. Clinicians in Basic Health Units performed less accurate risk assessments compared with those in hospitals. CONCLUSIONS: This study highlights important discrepancies between clinicians' management of human cases potentially exposed to rabies and recommendations in the NRMG. In particular, clinicians were not accurately assessing rabies risk in potentially exposed cases and were not basing their PEP treatment on the basis of their risk assessment. This has significant implications for achieving the national goal of eliminating dog-mediated human rabies by 2030 and may result in unnecessary costs associated with PEP. Recommendations to improve clinician's management of human cases potentially exposed to rabies include: reviewing and updating the NRMG, providing clinicians with regular and appropriately targeted training about rabies risk assessment and PEP prescription, and regularly reviewing clinicians' practices.

SciQuest and the complete archive of the NZVJ launched online.

The complete archive of the New Zealand Veterinary Journal went online on 1 January 2003 on SciQuest (www.sciquest.org.nz), an innovative new e-publishing website developed by the New Zealand Veterinary Association (NZVA). This follows hot on the heels of the CD-Rom version released from the same platform in June last year. Subscribers in 2003 will be able to search and retrieve, online, the full text of any article published in the journal since its inception in 1952. To date, few veterinary science journals are able to offer such a complete and comprehensive resource. SciQuest's mission is to provide high-speed access to science and continuing education articles to scientists and veterinarians in a fully indexed and searchable format, both online (via the internet) and offline (via CD-Rom). Searching and browsing the archive online is free and abstracts are available for all articles published from 1972 onwards. Access to full text, which is presented in PDF format exactly as published, is restricted to journal subscribers. The site will be updated with each issue of the journal as it is published. The online format will largely suit institutional users and casual enquiries, whereas the offline version is designed to be loaded onto computer hard drives for immediate access, any-time, and is better suited to frequent users and veterinary practitioners. A major advantage of the offline resource is its speed and immediacy, returning search results and full text much faster than the online version, without the need for internet access. The search engine has been designed to suit specific problem-oriented enquiries, and great effort has been expended to ensure the archive is both complete and accurate. The same search engine and interface has been provided on both versions, which allows for fast and highly specific searching of titles and full text of all articles for the occurrence of any word or combinations of words, as well as indexes of author, volume, year, article type, animal type and subject. Search results can be instantly sorted by author and date of publication, and powerful secondary search features such as 'by the same author', 'in the same issue' and 'same subject' links add power and flexibility. The offline version will be updated with annual releases and is available for purchase to subscribers and members of the NZVA.

Re: Journal Standards - Editor's reply.

Dr Jackson's concerns about citing non-peer-reviewed studies in the New Zealand Veterinary Journal (NZVJ) are shared by the editors and Editorial Board and have provoked considerable discussion about our policy and practice in this regard. Independent peer review remains a central part of the quality control process applied to the publication of scientific literature and is embraced by the NZVJ. All papers published in the NZVJ are scrutinised by the editors and Editorial Board, and those considered suitable are sent to two or three expert referees. In 2002, these comprised 195 people, of whom 36% were resident overseas. In addition, each paper is allocated a Guardian, a member of the Editorial Board specifically responsible for ensuring the adequacy of the peer-review process and providing an additional perspective regarding the quality of the paper. Papers are only accepted for publication after authors have satisfactorily addressed the issues raised by referees, Guardians and the Editor and sometimes by the Editorial Board. Thus, in addition to peer review, key quality-control roles are exercised at multiple points in the publication process by the Editor and Editorial Board to ensure the effectiveness, impartiality and integrity of the peer-review and editorial processes. Citation of data published in non-peer-reviewed sources in the NZVJ is discouraged but not precluded, in recognition of publications such as Surveillance and annual conference proceedings of New Zealand Veterinary Association (NZVA) special interest branches and the New Zealand Society of Animal Production as first repositories of disease reports and trial data often of substantial value and relevance to the scientific community. Many do not progress into the peer reviewed literature for reasons other than lack of quality and would otherwise be very difficult to access. As Dr Jackson points out, a substantial body of data generated in the deer industry has not yet appeared in peer-reviewed scientific journals, yet circulates widely in that industry. Considerable discussion preceded the permission granted authors to cite non-peer-reviewed studies to the extent evident in the December 2002 issue which was dedicated to feature review articles on the health and welfare of farmed deer. The proceedings of the annual seminars of the Deer Branch of the NZVA contain one of the most comprehensive records of studies on the nutrition, health and welfare of farmed deer available in the world. It was considered that the benefits of including discussion of these studies and exposing them to critical review outweighed the risks elaborated by Dr Jackson. However, the concerns raised by Dr Jackson and others prompted the Editorial Board to review the NZVJ's policy in this regard, which is now as follows: - Authors are specifically encouraged to cite peer-reviewed references wherever possible and particularly discouraged from citing their own publications in non-peer-reviewed sources. - Referees are specifically requested to scrutinise the use of non peer-reviewed references and make recommendations regarding alternatives or omission. - The Editor requests that authors: justify the use of non-peer-reviewed references; avoid the use of non-peer-reviewed citations more than 2 years old and; clearly distinguish references to nonpeer-reviewed references in the text of their papers, using qualifying statements such as preliminary data or non-peer-reviewed reportsetc. By limiting but not precluding citation of non-peer-reviewed studies, exposing them to specific scrutiny during the peer-review and editorial processes and by distinguishing them from peer-reviewed sources in the body of articles we aim to minimise the dangers of appearing to legitimise non-peer-reviewed data without losing the information value contained in some of these reports. There are many benefits of publishing in quality peer-reviewed journals such as the NZVJ, not least of which is the almost inevitable improvement in the quality and rigour of manuscripts after peer-review and the greatly increased opportunity for recognition and citation of peer-reviewed studies by others. Peer-reviewed journals remain the most widely accepted and established medium for communication and preservation of high-quality science. Authors and industry sectors that choose not to publish in peer-reviewed journals arguably limit the recognition and true progress of their science.

Gonadotrophin responsiveness, aromatase activity and insulin-like growth factor binding protein content of bovine ovarian follicles during the first follicular wave.

The aim of this study was to examine the function of granulosa cells and hormone concentrations in follicular fluid in bovine ovarian follicles during selection of the first dominant follicle. Ovaries were obtained from beef heifers on days 1-5 after ovulation: follicles > 4 mm in diameter were dissected and follicular fluid and granulosa cells were collected from individual follicles. Oestradiol production by granulosa cells after culture with testosterone was used to determine aromatase activity and responsiveness to gonadotrophins was determined by cAMP production after culture with FSH or LH. Concentrations of oestradiol, progesterone and insulin-like growth factor binding proteins (IGFBPs)-4 and -5 were measured in follicular fluid. Follicles were classified as largest or smaller (days 1 and 2), or dominant or subordinate (days 3-5). Aromatase activity was greater in granulosa cells from the largest follicle than in granulosa cells from smaller follicles on days 1, 3, 4 and 5 (P < 0.05). Responsiveness to LH was not detected in granulosa cells on day 1, but from day 2 to day 5 cells from the largest follicle were significantly more responsive than cells from smaller follicles (P < 0.05). Responsiveness to FSH was detected in granulosa cells from all follicles from day 1 onwards and did not differ between cells from the largest follicle or smaller follicles on any day. Follicular fluid concentrations of oestradiol and the ratio of oestradiol:progesterone were greater and concentrations of IGFBP-4 and -5 were lower in the largest follicle than in smaller follicles from day 2 to day 5 (P < 0.05). In conclusion, selection of the dominant follicle is associated with increased granulosa cell aromatase activity followed by increased cAMP response to LH and follicular fluid oestradiol concentrations, and decreased follicular fluid concentrations of IGFBP-4 and -5 within 2 days after ovulation.

Treatment of noncyclic lactating dairy cows with progesterone and estradiol or with progesterone, GnRH, prostaglandin F2 alpha, and estradiol.

The efficacy of two programs for treating noncyclic cows was compared. In trial 1, 478 cows in five herds were randomly divided into two groups. Cows in one group (C group) were treated with an intravaginal progesterone device for 8 d followed in 48 h by 1 mg of estradiol benzoate to cows that had not been detected in estrus since device removal. Those in the other group (CGP group) were treated with progesterone and estradiol as for the C group plus 10 micrograms of a GnRH agonist (buserelin) at device insertion and 25 mg of PGF2 alpha 7 d after device insertion. In trial 2 with 729 cows in nine herds, the treatments were similar to those in trial 1 except that the duration of progesterone treatment was 7 d. No significant difference was found between trials and results from both trials were combined. Compared with C group cows, CGP group cows had a greater estrous response rate (93.2 vs. 89.1%), a greater conception rate to first artificial insemination (AI, 47.1 vs. 29.4%), marginally lower conception rate to second AI (52.9 vs. 59.7%), lower nonpregnancy rate (8.3 vs. 11.1%), and shorter intervals from the start of breeding to conception by AI (9.8 vs. 15.3 d) or by AI or natural mating (21.6 vs. 26.3 d). The treatment protocol used for the CGP group achieved better reproductive performance than that used for the C group.

Granulosa cell apoptosis, aromatase activity, cyclic adenosine 3',5'-monophosphate response to gonadotropins, and follicular fluid steroid levels during spontaneous and induced follicular atresia in ewes.

The aims of the present study in ewes were 1) to test the hypothesis that apoptosis in granulosa cells is one of the processes involved in the structural demise of follicles and 2) to define the temporal relationships among the occurrence and degree of apoptosis in granulosa cells, aromatase activity, production of cyclic AMP (cAMP) by granulosa cells in response to FSH or LH, concentrations of estradiol 17 beta (E2) and progesterone in follicular fluid, and the characteristic morphometric changes associated with the process of follicular atresia. To address these aims, ewes were treated with either saline or steroid-free bovine follicular fluid (bFF) at 60 h after estrus, and ovarian follicles > or = 3 mm diameter were recovered at 0, 12, 18, or 24 h later. Apoptotic granulosa cells were identified by the presence of oligonucleosomes after 3'-end labeling of extracted DNA with [32P]alpha dideoxy ATP (ddATP). The degree of oligonucleosome formation, based on the intensity of radiolabeling, was given an apoptosis score (AP) of 0 (nondetectable), 1 (slight), 2 (moderate), or 3 (marked). Moreover, a labeling index (LI) was calculated from the amount of radiolabeled ddATP incorporated into low-molecular weight (< 4.2 kb) DNA fragments. On the basis of morphometric criteria, 73% (141 of 194) of the follicles classified as healthy had apoptotic granulosa cells compared to 86% (18 of 21) of the follicles classified as atretic. In the bFF-but not saline-treated ewes, the concentrations of plasma FSH had declined to basal values at 12 h after treatment. At the beginning of the treatment period, the degree of granulosa cell apoptosis was either undetectable (AP = 0, 47% of follicles) or slight (AP = 1, 44% of follicles) in the majority of follicles. After 12 h from the bFF but not the saline injection, there was a significant increase in the proportion of follicles (> or = 3 mm diameter) per ewe containing apoptotic granulosa cells (p < 0.001) and a significant decrease in the number of follicles per ewe with aromatase activity (p < 0.05) and with follicular fluid E2 > 20 ng/ml (p < 0.05). By 24 h after bFF treatment, apoptosis was evident in all follicles (> or = 3 mm diameter), fewer follicles contained FSH-responsive granulosa cells in terms of cAMP production (p < 0.05), and none were LH-responsive. A significant negative relationship was found between the degree of granulosa cell death as measured by L1 and follicular fluid E2 concentrations. In summary, the presence of apoptotic granulosa cells in an appreciable number of follicles considered to be healthy by morphometric criteria and before their commitment to preovulatory enlargement and ovulation suggests that apoptosis may be a physiological process in developing follicles and/or a very early event in atresia. Collectively, these data provide strong evidence that granulosa cells may die by apoptosis before there is an appreciable decrease in the capacity of the granulosa cell layer as a whole to respond to gonadotropins or to produce E2.

Morphological evidence of apoptosis and the prevalence of apoptotic versus mitotic cells in the membrana granulosa of ovarian follicles during spontaneous and induced atresia in ewes.

Apoptosis is a process by which granulosa cells are thought to be deleted during ovarian follicular atresia. The aims of the present studies, using sheep as the experimental model, were to determine 1) whether morphological changes in cells composing the membrana granulosa during the process of atresia conformed with the general criteria of apoptotic cell death as assessed using tissue sections stained with hematoxylin and eosin; 2) whether cells classified as apoptotic on the basis of their morphology contained fragmented DNA using an in situ 3' end-labeling technique; and 3) the degree of apoptosis and mitosis within the granulosa cell populations of large antral follicles (> or = 3 mm in diameter) during both spontaneous and experimentally induced atresia using stereological methods. The results showed that most degenerate granulosa cells in follicles undergoing atresia display the morphological characteristics of apoptosis, suggesting that this is the most common pathway of cell deletion. Typical features were cells containing nuclei with marginated chromatin; cells with a single small densely staining nucleus (pyknotic appearance); cells with multiple smaller, densely staining nuclear fragments; and densely staining membrane-bound bodies (apoptotic bodies) either singly or in clusters. Cells with morphological features more typical of oncosis or necrosis were sometimes observed, but mainly during the later stages of atresia. All cells classified as apoptotic on the basis of morphological criteria contained fragmented DNA as measured by 3' end-labeling. Apoptotic bodies and/or cells were found in all follicles examined, including those classified as healthy. The overall prevalence of apoptotic cells plus apoptotic bodies expressed as a percentage of the total granulosa cell number per follicle varied from 0.02% to 0.20% in healthy follicles, varied from 0.21% to 2.00% in follicles in early (primary) atresia, and was > 2.0% in follicles in later (secondary) atresia. Percentages of mitotic cells in healthy follicles were > 0.5% in all but one of those examined and were < 1.0% in all follicles classified as atretic. Both morphological and 3' end-labeling results indicated that apoptotic cells were widely disseminated throughout the membrana granulosa, including the cell layer adjacent to the basement membrane. Collectively, these observations indicate that during early atresia, apoptosis occurs randomly and is not limited to specific areas within follicles. Our finding that apoptotic cell death and mitosis occur simultaneously within the same follicle is consistent with the notion that atresia is determined by a dynamic equilibrium between cell division, differentiation, and death.

Increased bax and interleukin-1beta-converting enzyme messenger ribonucleic acid levels coincide with apoptosis in the bovine corpus luteum during structural regression.

Recent reports have demonstrated that prostaglandin F2alpha-induced generation of reactive oxygen species or their intermediates inhibits progesterone synthesis and may also serve as a trigger for apoptosis in the corpus luteum (CL). BCL-2, an inhibitor of apoptosis in a wide variety of cell types, has been reported to prevent oxidative stress-induced cell death. Thus, the present studies were conducted to determine whether levels of mRNA encoding BCL-2 and related members of this gene family (BAX and BCL-Xshort, which induce apoptosis; BCL-Xlong, a BCL-2 homologue that prevents apoptosis) differed in functional (Day 21 of pregnancy) versus regressed (Day 21 of the estrous cycle) CL in the bovine ovary. Levels of mRNAs encoding p53, a transcriptional regulator of the bcl-2 and bax genes, and interleukin-1beta-converting enzyme (ICE), a protein recently implicated in the induction of apoptosis whose expression may be enhanced by oxidative stress, were also assessed. Partial cDNA clones encoding bovine bax, bcl-x, p53, and Ice were isolated using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique with total RNA prepared from functional or regressed CL. A bovine bcl-2 cDNA could not be isolated from luteal tissue RNA despite the use of several primer pairs for amplification. Total RNA was then extracted from functional or regressed CL and analyzed by Northern blot analysis. The occurrence of apoptosis in regressed CL, as evidenced by the presence of internucleosomal DNA cleavage, was associated with a significant increase in both bax and Ice mRNA levels as compared with levels of bax and Ice expression in functional CL (p < 0.05, n = 3). There were no significant differences in bcl-x or p53 mRNA levels in functional versus regressed CL. Analysis of bcl-x mRNA by RT-PCR revealed that the long form was the primary, if not only, mRNA expressed in functional and regressed bovine luteal tissue. On the basis of data that increased expression of bax is associated with, and may be required for, apoptosis in ovarian granulosa cells and germ cells, we propose that BAX may play a similar role in apoptosis induction during luteal regression. Moreover, the increased Ice mRNA levels in regressed CL provides the first evidence that the ICE family of death proteases may be involved in luteolysis.

Physiological effects of undernutrition on postpartum anoestrus in cows.

The focus of this review is the physiological effects of undernutrition on ovarian follicle growth and the occurrence of ovulation in postpartum cows. Evidence suggests that moderate levels of underfeeding, before or after calving, may interfere with the mechanism(s) of final follicle maturation and ovulation, whereas more pronounced nutritional deficiencies may affect the mechanism(s) regulating dominant follicle size and the dynamics of dominant follicle growth and regression. Severe undernutrition may result in the absence of follicles > 5 mm in diameter. These changes are consistent with likely effects of reduced LH or FSH secretion that have been associated with inhibition of both tonic and surge release of GnRH from the hypothalamus. As yet, there is no direct evidence for local effects of undernutrition on ovarian function in cows, and studies examining this tissue often ignore dynamic interdependencies between the hypothalamus, anterior pituitary gland, and the ovaries, and possible nutritional modulation of ovarian feedback mechanisms that affect gonadotrophin secretion. Further research is required to elucidate nutritional effects on the positive feedback regulation of gonadotrophin secretion and on the expression of behavioural oestrus. Further studies are also needed to determine whether nutritional or metabolic state may influence ovarian function directly in a manner that affects the duration of postpartum anoestrus and fertility in postpartum cows.

Apoptosis in bovine granulosa cells in relation to steroid synthesis, cyclic adenosine 3',5'-monophosphate response to follicle-stimulating hormone and luteinizing hormone, and follicular atresia.

Apoptosis is a process of selective cell deletion implicated as a mechanism underlying the process of ovarian follicular atresia. The aims of this study were 1) to test the hypothesis that granulosa cell death during follicular (> or = 4 mm diameter) atresia in cows occurs by apoptosis and 2) to define relationships between the occurrence and degree of granulosa cell apoptosis, cAMP response to FSH or LH, extant aromatase activity, and other previously established biochemical and morphometric indices of granulosa cell function and follicular atresia in this species. Granulosa cells and follicular fluid from individual follicles 4-18 mm in diameter were collected from luteal-phase cow ovaries. Follicles were classified by morphometric criteria as "healthy" (n = 45) or atretic (n = 34). Apoptosis in granulosa cells from each follicle was inferred from detection of internucleosomal DNA cleavage by 3'-end radiolabeling; it was quantitated both subjectively from intensity of oligonucleosome labeling (apoptosis [AP] score = 0, 1, 2, or 3) and objectively by beta-counting of low-molecular weight gel fragments (labeling index; LI). Extant aromatase activity (ng estradiol produced/10(6) cells/3 h) and cAMP response (pmol/10(6) cells) to different doses of FSH or LH (1-10,000 ng/ml) was determined for granulosa cells from most healthy follicles (n = 39). Apoptosis was detected in granulosa cells from all atretic follicles as well as from 76% of healthy follicles, from 80% (16 of 20) of follicles with follicular fluid estradiol to progesterone ratios > 1, and from 71% (10 of 14) of follicles with extant aromatase activity (> 2 ng/10(6) cells/3 h). For healthy and atretic follicles, degree of DNA fragmentation was inversely related to the number of granulosa cells recovered (as percentage maximum by follicle size). In healthy follicles, FSH stimulated cAMP synthesis is a dose-dependent manner in granulosa cells from all follicles examined (> or = 4 mm), but only 36% of these had appreciable aromatase activity. The cAMP response to FSH (per cell) increased with follicle size from 4-7 mm in diameter and remained high in granulosa cells from follicles > or = 8 mm with aromatase activity; in cells without aromatase activity, cAMP response to FSH decreased with increasing follicle size > or 8 mm. The cAMP response to LH was generally low or undetectable in granulosa cells from 4-8-mm follicles; it then increased linearly with increasing follicle diameter > or = 8 mm, but to a greater degree in cells with aromatase activity than in cells without.(ABSTRACT TRUNCATED AT 400 WORDS)

Viruses associated with respiratory disease of horses in New Zealand: an update.

Viruses causing or associated with respiratory disease in horses worldwide are reviewed. Results are presented from a serological survey of 121 New Zealand foals and horses that had been affected by respiratory disease, determining the prevalence of antibodies in this country to the major viruses associated with similar disease overseas. To date there is no evidence of equine influenza virus in New Zealand. Both equine herpesvirus type 1 and 2 have been frequently isolated and show high serological prevalences. Serological evidence of equine rhinovirus type 1 and type 2 is presented with a prevalence of 12.3% and 41.2% respectively observed in foal sera, and 37.7% and 84.9% in adult horse sera. Antibody reacting to equine viral arteritis virus antigen was detected in 3/121 test sera. Equine adenovirus has been isolated on occasions and has shown a 39% serological prevalence in one study reviewed. Progress in New Zealand equine virus research is discussed.