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Development and evaluation of indirect enzyme-linked immunosorbent assays for the determination of immune response to multiple clostridial antigens in vaccinated captive bred southern white rhinoceros (Ceratotherium simum simum).

ABSTRACT: BACKGROUND:An overall increase in poaching of white rhinoceros results in captive breeding becoming a significant component of white rhinoceros conservation. However, this type of conservation comes with its own difficulties. When wildlife is captured, transported and/or confined to a boma environment, they are more predisposed to diseases caused by bacterial organisms such as spore forming Clostridium spp. A southern white rhinoceros (Ceratotherium simum simum) population on a captive bred farm was suspected to be affected by Clostridium infections. These endangered animals were apparently exposed to Clostridium spp., in the conservation area previously used for cattle farming. The rhinoceros population on the breeding operation property was vaccinated with a multi-component clostridial vaccine registered for use in cattle. Multiple indirect enzyme-linked immunosorbent assays (iELISAs) were developed in order to evaluate the serum antibody titres of these vaccinated animals. In evaluating vaccine efficacy, the gold standard mouse neutralization test (MNT) was not available and therefore iELISAs were developed for the detection of serum antibodies to C. perfringens type A (alpha toxin), C. chauvoei (whole cell), C. novyi (alpha toxin), C. septicum (alpha toxin) and C. sordellii (lethal toxin) in the white rhinoceros population using international reference sera of equine origin. Antibody titres against each clostridial antigen was evaluated in the vaccinated white rhinoceros population (n?=?75). Analytical specificity showed slight cross-reactions for C. chauvoei and C. perfringens type A with the other antigens. Individual assay cut-off values were calculated with 95% confidence. Coefficient of variance (CV) values for both the international reference sera and in-house control sera across all the antigens were well below 16%, indicating good assay repeatability. This convenient and fast assay is suitable for monitoring humoral immune responses to clostridial antigens in vaccinated white rhinoceroses. RESULTS:Checkerboard titrations indicated optimal antigen and antibody concentrations to be used for each respective iELISA developed. Each titration set of the respective international reference and in-house control sera showed good repeatability with low standard deviations and coefficient of variance values calculated between repeats for each antigen. Individual assays proved repeatable and showed good analytical sensitivity and specificity. CONCLUSIONS:The developed iELISAs are able to evaluate antibody profiles of phospholipase C, C. chauvoei whole cells, TcnA, ATX, TcsL in white rhinoceros serum using international reference sera.

SUBMITTER: Buys A

PROVIDER: S-EPMC7541252 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Development and evaluation of indirect enzyme-linked immunosorbent assays for the determination of immune response to multiple clostridial antigens in vaccinated captive bred southern white rhinoceros (Ceratotherium simum simum).

Buys Angela A Crafford Jannie J van Heerden Henriette H

Acta veterinaria Scandinavica 20201007 1

<h4>Background</h4>An overall increase in poaching of white rhinoceros results in captive breeding becoming a significant component of white rhinoceros conservation. However, this type of conservation comes with its own difficulties. When wildlife is captured, transported and/or confined to a boma environment, they are more predisposed to diseases caused by bacterial organisms such as spore forming Clostridium spp. A southern white rhinoceros (Ceratotherium simum simum) population on a captive b ...[more]

PMID: 33028379

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Project description:All species in the extant Rhinocerotidae family are experiencing increased threats in the wild, making captive populations essential genetic reservoirs for species survival. However, managed species face distinct challenges in captivity, resulting in populations that are not self-sustaining. Captive southern white rhinoceros (Ceratotherium simum simum) have low reproductive rates and presumed acyclicity is common among females. Although many females fail to ovulate, follicle growth may occur and ovulation can be hormonally induced. Female southern white rhino (n = 6), housed as a bachelorette group, were determined to be ovulatory (n = 1) or anovulatory (n = 5) by serial ultrasound and fecal progestagen analysis. When follicles reached pre-ovulatory size (~35 mm), females (n = 4) were induced to ovulate in 11 trials with a GnRH analog (4.5 mg, SucroMate™) via single intramuscular injection. Nine trials resulted in ovulation (81.8%), all between 36 and 48 hours post-treatment. Ovulations were confirmed by progestagen elevation above baseline coincident with visualization of a corpus luteum (CL). Luteal phases were characterized as short (<50 days) or long (≥50 days). Between short and long cycles, only the number of days of progestagen above baseline was significantly different (P < 0.05), while days with visible luteal structures was not significant (P = 0.11). Both cycle types were observed following both spontaneous and induced ovulations. Furthermore, we showed that longer cycle lengths do not necessarily indicate early pregnancy loss as none of the females were bred or inseminated during the study. While anovulation is common in the southern white rhino captive population, ovulation induction can be achieved efficiently and predictably for use in conjunction with artificial insemination or to facilitate natural breeding. This information will lead to more efficient use of assisted reproductive technologies to overcome reproductive challenges in this species and to generate genetically healthy captive populations as a hedge against extinction.

Project description:Gut microbiota are essential to nutrient metabolism and the maintenance of hindgut health. The characterization of faecal bacterial communities from healthy individuals is important for the establishment of baseline data that can be compared to periods of gut dysbiosis. Diet is a key determinant of the faecal microbial community structure and generation of volatile fatty acids, a main energy source for the host. While rhinoceroses are herbivores, black rhinoceroses are browsers and white rhinoceroses are grazers. The objective of our study was to characterize and compare diets, faecal bacterial communities, nutrients and metabolites between and amongst Southern white rhinoceroses and Southern black rhinoceroses (n = 3 rhinos/species) managed at Disney's Animal Kingdom® . Faecal bacterial communities were similar between individual white rhinos and dissimilar between species and individual black rhinos. Faecal butyrate and propionate molar proportions and concentrations were greater in black rhinos than white rhinos, whereas lactate was greater in white rhinos. The Shannon diversity, total operational taxonomic units, and relative abundance of Firmicutes were greater in white than black rhinos. The relative abundance of Proteobacteria in faeces from black rhinos was 3-fold greater than from white rhinos. One black rhino had a greater relative abundance of Verrucomicrobia (7.45 ± 1.31%) than all other individual rhinos (0.01-1.37%). White rhinoceroses demonstrated similar abundances of bacterial phyla and communities between one another and by individual, while black rhinoceroses were more dissimilar by individual. The dissimilarities between black rhinos were suspected to be due to total diet consumption variability, including browse diversity, and lack of direct contact. In contrast, the white rhinos commingled (i.e. nose-to-nose contact) and consumed similar amounts of hay, pellets and training items. These results suggest that species-specific diets and the individual contribute to differences in faecal bacterial communities, nutrients and metabolites between black and white rhinos housed at the same institution.

Project description:Land mammals support and move their body using their musculoskeletal system. Their musculature usually presents varying adaptations with body mass or mode of locomotion. Rhinocerotidae is an interesting clade in this regard, as they are heavy animals potentially reaching three tons but are still capable of adopting a galloping gait. However, their musculature has been poorly studied. Here we report the dissection of both forelimb and hindlimb of one neonate and one adult each for two species of rhinoceroses, the Indian rhinoceros (Rhinoceros unicornis) and the white rhinoceros (Ceratotherium simum). We show that their muscular organisation is similar to that of their relatives, equids and tapirs, and that few evolutionary convergences with other heavy mammals (e.g. elephants and hippopotamuses) are present. Nevertheless, they show clear adaptations to their large body mass, such as more distal insertions for the protractor and adductor muscles of the limbs, giving them longer lever arms. The quantitative architecture of rhino muscles is again reminiscent of that of horses and tapirs, although contrary to horses, the forelimb is much stronger than the hindlimb, which is likely due to its great role in body mass support. Muscles involved mainly in counteracting gravity (e.g. serratus ventralis thoracis, infraspinatus, gastrocnemius, flexores digitorum) are usually highly pennate with short fascicles facilitating strong joint extension. Muscles involved in propulsion (e.g. gluteal muscles, gluteobiceps, quadriceps femoris) seem to represent a compromise between a high maximal isometric force and long fascicles, allowing a reasonably fast and wide working range. Neonates present higher normalized maximal isometric force than the adults for almost every muscle, except sometimes for the extensor and propulsor muscles, which presumably acquire their great force-generating capacity during the growth of the animal. Our study clarifies the way the muscles of animals of cursorial ancestry can adapt to support a greater body mass and calls for further investigations in other clades of large body mass.

Dataset Information

Development and evaluation of indirect enzyme-linked immunosorbent assays for the determination of immune response to multiple clostridial antigens in vaccinated captive bred southern white rhinoceros (Ceratotherium simum simum).

Publications

Development and evaluation of indirect enzyme-linked immunosorbent assays for the determination of immune response to multiple clostridial antigens in vaccinated captive bred southern white rhinoceros (Ceratotherium simum simum).

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