In Vitro Maturation of Dromedary (Camelus dromedarius) Oocytes: Effect of Different Protein Supplementations and Epidermal Growth Factor*

The present experiment was aimed to compare the effect of different protein supplementation sources, foetal calf serum (FCS), oestrous dromedary serum (EDS) and BSA, in experiment 1, and the effect of different concentrations of epidermal growth factor (EGF), in experiment 2, on in vitro nuclear maturation of the dromedary oocytes. Cumulus oocyte complexes (COCs) were harvested from the ovaries collected from a local slaughterhouse by aspirating the visible follicles in PBS supplemented with 5% FCS. Pooled COCs were randomly distributed to 4‐well culture plates containing 500 μl of the maturation medium and cultured at 38.5°C in an atmosphere of 5% CO₂ in air for 32–36 h. The basic maturation medium consisted of TCM‐199 supplemented with 0.1 mg/ml L‐glutamine, 0.8 mg/ml sodium bicarbonate, 0.25 mg/ml pyruvate, 50 μg/ml gentamicin, 10 μg/ml bFSH, 10 μg/ml bLH and 1 μg/ml estradiol. In experiment 1, this medium was supplemented with 10% FCS, 10% EDS or 0.4% BSA, whereas in experiment 2, it was supplemented with 0.4% BSA and 0, 10, 20 or 50 ng/ml of EGF. The oocytes were fixed, stained with 1% aceto‐orcein stain and their nuclear status was evaluated. Oocytes were classified as germinal vesicle, diakinesis, metaphase‐I, anaphase‐I (A‐I), metaphase‐II (M‐II) and those with degenerated, fragmented, scattered, activated or without visible chromatin as others. There was no difference (p > 0.05) observed in the proportion of oocytes reaching M‐II stage between the media supplemented with FCS (71.5 ± 4.8), EDS (72.8 ± 2.9) and BSA (72.7 ± 6.2). In experiment 2, a higher proportion (p < 0.05) of oocytes reached M‐II stage when the medium was supplemented with 20 ng/ml of EGF (81.4 ± 3.2) when compared with the media supplemented with 10 ng/ml (66.9 ± 4.1) and control (67.2 ± 7.1) groups. It may be concluded that the maturation media for dromedary camel oocytes can be supplemented with any of the three protein sources, i.e. FCS, EDS and BSA without any significant differences on the maturation rates. Also, a supplementation of 20 ng/ml of EGF in the maturation medium seems to be optimal and improves the nuclear maturation of dromedary camel oocytes.     

Camelid Immunoglobulins and Their Importance for the New‐Born – A Review

Camelid immunoglobulins differ from all other known antibodies and contradict all common theories on antibody diversity. It was demonstrated that up to 75 % of all serum proteins are immunoglobulin G (IgG) molecules lacking light chains. IgG₂ and IgG₃, which only consist of heavy chains, have a low molecular weight which improves their biodistribution and allows a better tissue penetration. Of special importance is the long complementary determining region (CDR) loop which inserts deep into the active site of an enzyme. This binding property was only observed in experiments to gain structural data and to point out the extraordinary value of heavy chain antibodies as biochemical and pharmacological tools. The acquisition and absorption of adequate amounts of colostral immunoglobulins are essential to the health of the neonate. Pre‐colostrum serum IgG levels in camelids are low, with concentrations of 0.26 ± 0.23 mg/ml. Maximum IgG levels are reached after 24 h and kept at a plateau with concentrations of 24.52 ± 8.8 mg/dl. IgG concentrations above 10 mg/ml indicate a successful passive transfer. IgG levels decline after 2–5 weeks and a marked increase is observed between 1 and 2 months, indicating that the immune system of the neonate has started to mature. A number of different tests are available for the assessment of IgG serum levels. Single radial immunodiffusion (SRID) is the only method that specifically measures serum IgG concentrations. It is a reliable assay to test failure of passive transfer (FPT). FPT is a major factor in neonatal mortality in camelids, but very little has been published so far. Therapeutic administration of colostrum will provide passive protection against infectious diseases for a 2–3‐week period of risk, and the intravenous administration of 20–40 ml of camelid plasma helps to combat FPT.     

Microarray-based genotyping of Staphylococcus aureus isolates from camels

Staphylococcus aureus is a common cause of mastitis and other diseases in camels. In order to obtain data on population structure as well as on the carriage of toxin genes and resistance markers, a collection of 45 isolates from dromedaries of Dubai, United Arab Emirates, were genotyped. These isolates belonged to clonal complexes CC6 (twenty isolates; 44.44%), CC30 (sixteen isolates; 35.56%), CC188 (five isolates; 11.11%), CC152 (1 isolate, 2.2%) and to a previously un-described sequence type (ST1755: arcc-18, aroe-115, glpf-6, gmk-2 pta-109, tpi-50 and yqil-2; three isolates; 6.67%). Resistance genes proved to be rare. Only three out of 45 isolates (6.67%) carried the beta-lactamase operon. The tetracycline resistance gene tetK was also detected in three isolates (6.67%). Neither the mecA gene, defining MRSA, nor other resistance genes were found. Common virulence markers included leukocidin genes lukD+lukE (in twenty-five isolates; 55.56%), the staphylokinase gene sak (twenty-two isolates; 48.89%), the enterotoxin gene cluster egc (fifteen isolates; 33.33%), and a distinct variant of the enterotoxin A gene (sea-320E, GenBank AY196686.1; thirteen isolates; 28.89%). One CC152 isolate was positive for genes encoding the Panton-Valentine leukocidin (lukF-PV+lukS-PV). This study provides first genotyping data on the population structure and the presence of toxin genes and resistance markers of S. aureus strains in Middle Eastern camels.     

Disseminated Rhodococcus equi infection in dromedary camels (Camelus dromedarius)

Rhodococcus (R). equi, a recognized pathogen in horses, is emerging as a human opportunistic pathogen, especially in immunocompromized people. It affects also New World camelids, but there are no reports of R. equi infection in Old World camelids yet. Four cases of disseminated R. equi infection in adult breeding dromedaries occurred at one camel farm near Dubai within 16 months of each other. At necropsy the lungs were diffusely consolidated with large caseous areas. Histology revealed severe suppurative to necrotising pneumonia with multiple encapsulated abscesses. Immunohistochemistry enabled the detection of 15- to 17-kDa antigens (VapA) of R. equi in the lung sections. High numbers of R. equi were isolated from the lung lesions as well as from liver, spleen and mediastinal lymph nodes, indicative of septicaemia. The isolated strains were PCR-positive for the specific virulence plasmid (VapA-Gen) of R. equi, indicating virulent strains and containing an 85-kb type I plasmid. This is the first report of disseminated R. equi infection in Old World camelids. Since adult camels in general do not suffer from bacterial caused pneumonia (except tuberculosis), this is a new emerging disease for camels.     

Is There a Two‐Humped Stage in the Embryonic Development of the Dromedary?

It has been postulated that the one‐humped (Arabian) dromedary and the two‐humped (Bactrian) camel originated from a single ancestor. Consequently, the dromedary was considered a breed of the two‐humped camel, based on an anatomical study by Lombardini L, 1879: Ann. Del. Universita Toscane, 259 , 147, who described a reduced second hump like structure in foetal and adult dromedaries. To resolve this lingering issue, we analysed dromedary foetuses and calves. In contrast to the situation in two‐humped camels, we never observed any rudimentary second hump in the dromedary foetuses or calves.     

Bacterial infertility in camels (Camelus dromedarius): isolation of Campylobacter fetus

This study constitutes the first reported isolation of C. fetus subsp. venerealis from cervical swabs in breeding camels. 9 of 30 camels which were barren for a long period were found to be positive for C. fetus. The methods of isolation are described. Additionally, cervical flora of 30 camels were examined. 9 different bacterial species were isolated of which Staphylococcus aureus was most frequently found. All samples were negative for Trichomonas sp. and Streptococcus zooepidemicus.     

Evaluation of a newly developed real-time PCR for the detection of Taylorella equigenitalis and discrimination from T. asinigenitalis

A 'culture-LightCycler PCR' assay has been developed for the detection of Taylorella equigenitalis, the causative agent of contagious equine metritis (CEM) in horses. The primers and hybridisation probes were derived from the 16S rDNA sequence. Their specificity was determined in two closely related organisms and six commensal bacteria of the genital tract. The assay was specific for T. equigenitalis and discriminates T. asinigenitalis isolates. It also avoids misidentifications of morphologically and phenotypically similar organisms. The sensitivity was evaluated in comparison to a standard bacteriological culture method. It detected T. equigenitalis in 10 of 52 samples that had not been identified bacteriologically. The results indicated that the assay had a greater sensitivity. This is the first real-time PCR for the detection of T. equigenitalis and avoids PCR carry-over contamination. The 'culture-LightCycler PCR' assay is specific, sensitive and reproducible, and can be used effectively for the detection of T. equigenitalis infections.     

Bumblefoot and lack of exercise among wild and captive-bred falcons tested in the United Arab Emirates

In the 1990s, bumblefoot posed a major health problem to the falcons in the United Arab Emirates. This retrospective study based on statistical field research showed that in captivity wild falcons need a training frequency of twice a day to reduce the bumblefoot morbidity rate. In this context, they responded very well to free flight in aviaries during the molting season. In contrast, captive-bred falcons did not show a highly significant difference regarding training frequencies. Wild falcons tested fed with pigeons, bustards, and ducks suffered significantly less from bumblefoot compared with those fed a diet of quail. Apart from the disease-reducing impact of beef and mice, captive-bred falcons tested did not show any significant difference regarding the influence of diet on the bumblefoot occurrence.