Isolation of camel single domain antibodies against Yersinia pestis V270 antigen based on a semi-synthetic single domain antibody library and development of a VHH-based lateral flow assay.

BACKGROUND: Antibodies have been proven effective as diagnostic agents for detecting zoonotic diseases. The variable domain of camel heavy chain antibody (VHH), as an antibody derivative, may be used as an alternative for traditional antibodies in existing immunodiagnostic reagents for detecting rapidly spreading infectious diseases. OBJECTIVES: To expedite the isolation of specific antibodies for diagnostic purposes, we constructed a semi-synthetic camel single domain antibody library based on the phage display technique platform (PDT) and verified the validity of this study. METHODS: The semi-synthetic single domain antibody sequences consist of two parts: one is the FR1-FR3 region amplified by RT-PCR from healthy camel peripheral blood lymphocytes (PBLs), and the other part is the CDR3-FR4 region synthesised as an oligonucleotide containing CDR3 randomised region. The two parts were fused by overlapping PCR, resulting in the rearranged variable domain of heavy-chain antibodies (VHHs). Y. pestis low-calcium response V protein (LcrV) is an optional biomarker to detect the Y. pestis infection. The semi-synthetic library herein was screened using recombinant (LcrV) as a target antigen. RESULTS: After four cycles of panning the library, four VHH binders targeting 1-270 aa residues of LcrV were isolated. The four VHH genes with unique sequences were recloned into an expression vector and expressed as VHH-hFc chimeric antibodies. The purified antibodies were identified and used to develop a lateral flow immunoassay (LFA) test strip using latex microspheres (LM) for the rapid and visual detection of Y. pestis infection. CONCLUSIONS: These data demonstrate the great potential of the semi-synthetic library for use in isolation of antigen-specific nanobodies and the isolated specific VHHs can be used in antigen-capture immunoassays.

Fetal gender prediction by monitoring circulating steroid hormones and some biochemical parameters in maternal dromedary camels.

To predict the sex of the foetus, healthy pregnant dromedary camels (n = 24) were included. Blood samples were collected for measurements of progesterone, estradiol, testosterone, and cortisol as well as total proteins, albumin, glucose, creatinine, blood urea nitrogen, phosphorus, calcium, creatine kinase, alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (GGT), calcium, phosphorus, and magnesium. Statistical analysis revealed differences between pregnant camels and pregnant camels in terms of female or male foetuses depending on the actual sex of the born calf. The results revealed that testosterone and ALP concentrations were significantly (P < 0.001) greater in camels given to males than in those given to calves. There were strong positive correlations between male calf birth and testosterone and ALP concentrations (r = 0.864; P < 0.0001 and r = 0.637; P < 0.001, respectively). On the other hand, the cortisol, glucose and creatinine concentrations were significantly lower (P lower in camel calved males than in females). There were significant negative correlations between male calf birth and the cortisol, glucose and creatinine concentrations (r =-0.401; P = 0.052; r =-0.445; P = 0.029 and r =-0.400; P = 0.053, respectively). The concentrations of calcium, phosphorus, calcium/phosphorus ratio, magnesium, and albumin and the albumin/globulin ratio were not significantly different (P > 0.05) between the two groups. In conclusion, testosterone could be used as a biomarker to determine the sex of foetuses in dromedary camels.

Genetic characterization of the prion protein gene in camels (Camelus) with comments on the evolutionary history of prion disease in Cetartiodactyla.

Transmissible spongiform encephalopathies (TSEs) are a fatal neurogenerative disease that include Creutzfeldt-Jakob disease in humans, scrapie in sheep and goats, bovine spongiform encephalopathy (BSE), and several others as well as the recently described camel prion disease (CPD). CPD originally was documented in 3.1% of camels examined during an antemortem slaughterhouse inspection in the Ouargla region of Algeria. Of three individuals confirmed for CPD, two were sequenced for the exon 3 of the prion protein gene (PRNP) and were identical to sequences previously reported for Camelus dromedarius. Given that other TSEs, such as BSE, are known to be capable of cross-species transmission and that there is household consumption of meat and milk from Camelus, regulations to ensure camel and human health should be a One Health priority in exporting countries. Although the interspecies transmissibility of CPD currently is unknown, genotypic characterization of Camelus PRNP may be used for predictability of predisposition and potential susceptibility to CPD. Herein, eight breeds of dromedary camels from a previous genetic (mitochondrial DNA and microsatellites) and morphological study were genotyped for PRNP and compared to genotypes from CPD-positive Algerian camels. Sequence data from PRNP indicated that Ethiopian camels possessed 100% sequence identity to CPD-positive camels from Algeria. In addition, the camel PRNP genotype is unique compared to other members of the Orders Cetartiodactyla and Perissodactyla and provides an in-depth phylogenetic analysis of families within Cetartiodactyla and Perissodactyla that was used to infer the evolutionary history of the PRNP gene.

Uterotubal junction of the bovine (Bos taurus) versus the dromedary camel (Camelus dromedarius): Histology and histomorphometry.

This study aims to conduct a comparative analysis of the uterotubal junction in two distinct mammalian species, the bovine (Bos taurus) and the dromedary camel (Camelus dromedarius), focusing on histological and histomorphometric parameters. Uterotubal junction (UTJ) was dissected from 8 cows and 12 camels with dominant follicles, and processed for H&E staining for histology, and histomorphometry examination. The results showed that the camel uterotubal junction papillae (UTJP) existed only in camels and was completely absent in cattle. Histologically, the cow UTJ appears with a star-shaped lumen, and the mucosa is lined by a simple columnar epithelium with ciliated and non-ciliated cells. Superficial (SG) and deep glands (DG) were abundant in the submucosa. Camel UTJP is a conical structure, it has a pale yellowish colour, 0.5 ± 0.2 cm height, and 0.3 ± 0.1 cm width, and it has a distinct sphincter at the tip towards the uterine lumen. In the camel UTJP, the lumen is wider, and the mucosa showed large multiple folds lined with ciliated and non-ciliated cells in a pseudostratified columnar epithelium. The submucosa showed no mucosal glands. Lumen area, lumen epithelial height, luminal epithelial density, the thickness of the muscular layer, number of folds, fold height, epithelial height, fold area and epithelial perimeter were higher in camel UTJP (p < .001) than cow UTJ, while the total endosalpinx area and glandular epithelial density were lower in camel UTJP (p < .001) than in cow UTJ. In conclusion, this study elucidates potential variations in the uterotubal junction between bovines and dromedary camels, providing valuable insights into their reproductive adaptations. The epithelial lining, absence of glands and the thick layer of tunica muscularis might indicate that camel UTJP could play a mechanical role in selecting spermatozoa and assisting the hatching of blastocysts during their passage into the uterus.

Epidemiology and diagnostic accuracy of Clostridium perfringens toxins in the intestinal contents of camels, sheep, and cattle: a cross-sectional study in Dakahlia governorate, Egypt.

This study aimed to establish an accurate epidemiological surveillance tool for the detection of different C. perfringens types from 76 diseased and 34 healthy animals in Dakhalia Governorate, Egypt. A total of 110 intestinal content samples were randomly collected from camels, sheep, and cattle. C. perfringens was isolated and biochemically identified by the VITEK2 system. Toxinotyping and genotyping of C. perfringens isolates were specified by a multiscreen ELISA and real-time qPCR (rt-qPCR). The occurrence of C. perfringens was highest among camels (20% in healthy and 25% in diseased) and was lowest in cattle (23.1% and 14.7%). The cpa toxin was detected in all isolates by rt-qPCR and in 7 isolates by ELISA, ext toxin was detected in 7 isolates by rt-qPCR and in 6 isolates by ELISA, and cpb toxin was detected in 2 isolates by both rt-qPCR and ELISA. Four types of C. perfringens were identified by rt-qPCR, type A (65.2%), B (4.3%), C (4.3%), and D (26.1%), and three types by ELISA, type D (17.4%), A (8.7%) and C (4.3%). Our study indicated the prevalence of infection in Dakahlia by C. perfringens type A and D, particularly camels, and recommends adopting an appropriate vaccination strategy among the studied animals.

A sero-epidemiological analysis of Coxiella burnetii infection and its risk factors in livestock from Addis Ababa, Adama, and Modjo abattoirs and pastoral areas of Oromia, Ethiopia.

BACKGROUND: Coxiella burnetii is causing infections in both humans and animals, resulting in Q fever and Coxiellosis, respectively. Information on the occurrence of C. burnetii infection is scarce in Ethiopia. This study estimated the sero-prevalence of C. burnetii infection and associated risk factors in four common livestock species from Addis Ababa, Adama, and Modjo abattoirs and pastoral areas of Oromia, Ethiopia. RESULTS/PRINCIPAL FINDINGS: Sera samples were analyzed for the presence of anti-C. burnetii antibodies using an indirect Enzyme Linked Immunosorbent Assay kit. Out of the 4140 serum samples tested, 777 (18.77%; 95% CI: 17.59, 19.99) were found positive for C. burnetii. The sero-prevalence estimate was 27.17% at Addis Ababa abattoir, 19.41% at Adama abattoir, 19.13% at Modjo abattoir and 12.1% in animals tested from pastoral areas. Sera analysis at the animal species level showed that cattle exhibited the lowest sero-prevalence estimate (11.83%; 95% CI, 10.27-13.53%), while the highest was observed in camels (28.39%; 95% CI, 25.16-31.80%). The sero-prevalence estimate was 21.34% (95% CI, 18.86-23.99%) in goats and 20.17% (95% CI, 17.49-23.07%) in sheep. The results of multivariable logistic regression analysis showed that species, age, sex of animals and tick infestation were important risk factors for C. burnetii infection. The odds of infection were 3.22 times higher in camels and almost twice as high in goats and sheep compared to cattle. Adult animals were infected more likely (OR = 3.23) than young ones. Interestingly, a significant difference was observed in the sero-prevalence of infection between animals that were infested with ticks (OR = 16.32) and those which were tick-free. CONCLUSION: This study provides valuable insights into the sero-epidemiology of C. burnetii infection in four common livestock species at major abattoirs and pastoral areas of Ethiopia. The findings highlight the need for further studies and implementing surveillance and biosecurity measures to prevent the spread of the disease in both humans and livestock to safeguard the economical and public health aspects.

Molecular identification and morphological variations of Amblyomma lepidum imported to Egypt, with notes about its potential distribution under climate change.

The tick Amblyomma lepidum is an ectoparasite of veterinary importance due to its role in transmitting livestock diseases in Africa, including heartwater. This study was conducted in 2023 to monitor Amblyomma spp. infestation in dromedary camels imported from Somalia, Ethiopia, and Sudan to Egypt. This study inspected 200 camels at the Giza governorate's camel market that had been imported from Somalia, 200 from Ethiopia, and 200 from Sudan for tick infestation. Specimens were identified using morphological characteristics and phylogenetic analyses of the 12S and 16S rRNA genes. Clusters were calculated using an unweighted pair-group method with arithmetic averages (UPGMA) dendrogram to group the specimens according to their morphometric characteristics. The morphometric analysis compared the body shape of ticks collected from different countries by analyzing dorsal features. Principal component analysis (PCA) and canonical variate analysis (CVA) were performed to obtain body shape variation among specimens from different countries. Results indicated that camels were infested by 57 males Amblyomma lepidum, and no female specimens were observed; among these specimens, one may have a morphological abnormality. The results suggest that A. lepidum specimens collected from camels imported to Egypt from African countries exhibit locally adapted morphology with variations among specimens, particularly variations in body size. This adaptation suggests minimal potential for genetic divergence. Ecological niche modeling was used to predict the areas in Africa with suitable climates for A. lepidum. The study confirmed that East African countries might have the most favorable climatic conditions for A. lepidum to thrive. Interestingly, the amount of rain during the wettest quarter (Bio16) had the strongest influence on the tick's potential distribution, with suitability decreasing sharply as rainfall increased. Future predictions indicate that the climatic habitat suitability for A. lepidum will decrease under changing climate conditions. However, historical, current, and future predictions indicate no suitable climatic habitats for A. lepidum in Egypt. These findings demand continuous surveillance of A. lepidum in camel populations and the development of targeted strategies to manage tick infestations and prevent the spread of heartwater disease.

Impact of quercetin conjugation using alkaline and free radical methods with tandem ultrasonication on the functional properties of camel whey and its hydrolysates.

The structural and functional properties of whey-quercetin and whey hydrolysate-quercetin conjugates synthesized using alkaline and free radical-mediated methods (AM and FRM) coupled with sonication were studied. FTIR showed new peaks at 3000-3500 cm-1 (N-H stretching regions) and the 1000-1100 cm-1 region with the conjugates. Conjugation increased the random coils and α-helix content while decreasing the ß-sheets and turns. It also increased the particle size and surface hydrophobicity which was significantly (p < 0.05) higher in AM than FRM conjugates. AM conjugates had higher radical scavenging activity but lower quercetin content than FRM conjugates. Overall, the functional properties of whey-quercetin conjugates were better than whey hydrolysate-quercetin conjugates. However, hydrolysate conjugates had significantly higher denaturation temperatures irrespective of the method of production. Sonication improved the radical scavenging activity and quercetin content of FRM conjugates while it decreased both for AM conjugates. This study suggested that whey-quercetin conjugates generally had better quality than whey hydrolysate conjugates and sonication tended to further improve these properties. This study highlights the potential for using camel whey or whey hydrolysate-quercetin conjugates to enhance the functional properties of food products in the food industry.

Serosurvey and associated risk factors for Chlamydia abortus infection in Dromedary camels in Egypt.

Chlamydia abortus (C. abortus) is a gram-negative, obligate intracellular bacterium that causes major public health problems in human and reproductive problems in animals. The information about the epidemiology of this pathogen among camels in Egypt is very rare. This study aimed to evaluate the existence of antibodies against C. abortus in camels and assess the related risk factors for infection. A total of 410 blood samples were collected from camels from three Egyptian governorates and examined using commercial ELISA kit. The overall seroprevalence rate was 6.6% and the higher C. abortus seropositivity rate was found in Giza governorate. Location, sex and infestation by ectoparasites did not influence on the seroprevalence of the disease. In addition, age, herd size, contact with small ruminants and history of abortion were identified as risk factors for C. abortus infection according to the univariate analysis. Based on multivariate analysis, age group of 4-8 years, small herd size, contact of camels with sheep and goats, and history of abortion were found to be significant risk factors for chlamydiosis transmission in camels. These factors had odds ratios of 4.23, 3.51, 2.84, and 2.5, respectively. These results suggest that camels have a role in the epidemiology of C. abortus infection. This promotes awareness and severe public health concern about infectious camel illnesses, allowing for additional diagnostic advancements and effective management techniques to be developed.

Seroprevalence of brucellosis and associated risk factors in camels and its herders in selected districts of Somali Pastoral Region, Eastern Ethiopia.

Brucellosis poses a major public and animal health problem in many parts of the world, particularly in pastoral settings, however, seroepidemological studies are scarce. A cross-sectional study was conducted from December 2021 to April 2022 to estimate the prevalence of brucellosis and to identify the associated risk factors for camels and occupational individuals from three purposively selected districts of the Somali pastoral region in Eastern Ethiopia. Serum samples were serially diluted using the Rose Bengal Plate Test (RBPT) as a screening test and a competitive Enzyme-Linked Immunosorbent Assay (cELISA) test as a confirmatory test. From a total of 450 camels and 250 human serum samples tested, the overall seroprevalence was confirmed to be 2.9 % (95 % CI, 1.5-4.9) in camels and 2.0 % (95 % CI, 0.2-3.7) in humans. In camels, abortion and retained fetal membrane (RFM) were significant risk factors for Brucella seropositivity (p<0.05). However, in humans, RFM disposal differed significantly (p<0.05). The fact that brucellosis is found in both camels and humans highlights the importance of implementing a coordinated One Health approach to control and eliminate the disease. This would ensure improved public health and increased livestock productivity.

Characterization of neutralizing chimeric heavy-chain antibodies against tetanus toxin.

Tetanus toxin (TeNT) is one of the most toxic proteins. Neutralizing antibodies against TeNT are effective in prevention and treatment. In this study, 14 anti-tetanus nanobodies were obtained from a phage display nanobody library by immunizing a camel with the C-terminal receptor-binding domain of TeNT (TeNT-Hc) as the antigen. After fusion with the human Fc fragment, 11 chimeric heavy-chain antibodies demonstrated nanomolar binding toward TeNT-Hc. The results of toxin neutralization experiments showed that T83-7, T83-8, and T83-13 completely protected mice against 20 × the median lethal dose (LD50) at a low concentration. The neutralizing potency of T83-7, T83-8, and T83-13 against TeNT is 0.4 IU/mg, 0.4 IU/mg and 0.2 IU/mg, respectively. In the prophylactic setting, we found that 5 mg/kg of T83-13 provided the mice with full protection from tetanus, even when they were injected 14 days before exposure to 20 × LD50 TeNT. T83-7 and T83-8 were less effective, being fully protective only when challenged 7 or 10 days before exposure, respectively. In the therapeutic setting, 12 h after exposure to TeNT, 1 ~ 5 mg/kg of T83-7, and T83-8 could provide complete protection for mice against 5 × LD50 TeNT, while 1 mg/kg T83-13 could provide complete protection 24 h after exposure to 5 × LD50 TeNT. Our results suggested that these antibodies represent prophylactic and therapeutic activities against TeNT in a mouse model. The T83-7, T83-8, and T83-13 could form the basis for the subsequent development of drugs to treat TeNT toxicity.

Structural basis for potent neutralization of human respirovirus type 3 by protective single-domain camelid antibodies.

Respirovirus 3 is a leading cause of severe acute respiratory infections in vulnerable human populations. Entry into host cells is facilitated by the attachment glycoprotein and the fusion glycoprotein (F). Because of its crucial role, F represents an attractive therapeutic target. Here, we identify 13 F-directed heavy-chain-only antibody fragments that neutralize recombinant respirovirus 3. High-resolution cryo-EM structures of antibody fragments bound to the prefusion conformation of F reveal three distinct, previously uncharacterized epitopes. All three antibody fragments bind quaternary epitopes on F, suggesting mechanisms for neutralization that may include stabilization of the prefusion conformation. Studies in cotton rats demonstrate the prophylactic efficacy of these antibody fragments in reducing viral load in the lungs and nasal passages. These data highlight the potential of heavy-chain-only antibody fragments as effective interventions against respirovirus 3 infection and identify neutralizing epitopes that can be targeted for therapeutic development.

Screening, expression and anti-tumor functional identification of anti-LAG-3 nanobodies.

OBJECTIVE: To screen and obtain specific anti-lymphocyte activation gene-3 (LAG3) nanobody sequences, purify and express recombinant anti-LAG3 nanobody, and verify its effect on promoting T cells to kill tumor cells. METHODS: Based on the camel derived natural nanobody phage display library constructed by the research group, the biotinylated LAG3 antigen was used as the target, and the anti-LAG3 nanobody sequences were screened by biotin-streptavidin liquid phase screening, phage-ELISA and sequencing. The sequence-conjµgated human IgG1 Fc fragment was obtained, the recombinant anti-LAG3 nanobody expression vector was constructed, the expression of the recombinant anti-LAG3 nanobody was induced by IPTG and purified, and the characteristics and functions of the recombinant anti-LAG3 nanobody were verified by SDS-PAGE, Western blot, cytotoxicity assay, etc. RESULTS: One anti-LAG3 nanobody sequence was successfully screened, and the corresponding recombinant anti-LAG3 nanobody-expressing bacteria were constructed. The results of SDS-PAGE, Western blot and cytotoxicity assay showed that the recombinant anti-LAG3 nanobody was successfully expressed, which was specific, and it could promote the killing ability of T cells against tumor cells, and the optimal concentration was 200 µg/mL. CONCLUSION: The recombinant anti-LAG3 nanobody screened and expressed has specific and auxiliary anti-tumor cell effects, which lays a foundation for its subsequent application.

Application of gelatin-based zinc oxide nanoparticles bionanocomposite coatings to control Listeria monocytogenes in Talaga cheese and camel meat during refrigerated storage.

Listeria monocytogenes is a concerning foodborne pathogen incriminated in soft cheese and meat-related outbreaks, highlighting the significance of applying alternative techniques to control its growth in food. In the current study, eco-friendly zinc oxide nanoparticles (ZnO-NPs) were synthesized using Rosmarinus officinalis, Punica granatum, and Origanum marjoram extracts individually. The antimicrobial efficacy of the prepared ZnO-NPs against L. monocytogenes was assessed using the agar well diffusion technique. Data indicated that ZnO-NPs prepared using Origanum marjoram were the most effective; therefore, they were used for the preparation of gelatin-based bionanocomposite coatings. Furthermore, the antimicrobial efficacy of the prepared gelatin-based bionanocomposite coatings containing eco-friendly ZnO-NPs was evaluated against L. monocytogenes in Talaga cheese (an Egyptian soft cheese) and camel meat during refrigerated storage at 4 ± 1 oC. Talaga cheese and camel meat were inoculated with L. monocytogenes, then coated with gelatin (G), gelatin with ZnO-NPs 1% (G/ZnO-NPs 1%), and gelatin with ZnO-NPs 2% (G/ZnO-NPs 2%). Microbiological examination showed that the G/ZnO-NPs 2% coating reduced L. monocytogenes count in the coated Talaga cheese and camel meat by 2.76 ± 0.19 and 2.36 ± 0.51 log CFU/g, respectively, by the end of the storage period. Moreover, G/ZnO-NPs coatings controlled pH changes, reduced water losses, and improved the sensory characteristics of Talaga cheese and camel meat, thereby extending their shelf life. The obtained results from this study indicate that the application of gelatin/ZnO-NPs 2% bionanocomposite coating could be used in the food industry to control L. monocytogenes growth, improve quality, and extend the shelf life of Talaga cheese and camel meat.

Impact of papain on the treatment of raw diluted dromedary semen.

A variety of parameters, including liquefaction and semen viscosity, affect the sperm's ability to travel and reach the egg for fertilization and conception. Given that the details behind the viscosity of the semen in male camels have not yet been fully clarified, the purpose of this study was to ascertain how the addition of papain affected the viscosity of fresh diluted camel semen. The study examined semen samples derived from camels that had distinct viscosities. Sperm motility, viability, abnormal sperm percentage, concentration, viscosity, morphometry, acrosome integrity and liquefaction were among the evaluations following 0, 5, 10, 20 or 30 min of incubation at 37°C with papain (0.004 mg/mL, 0.04 mg/mL or 0.4 mg/mL; a semen sample without papain was used as a control). A statistically significant interaction between the effects of papain concentrations and incubation time was found (F = 41.68, p = .0001). Papain concentrations (p = .0001) and incubation times (p = .0001) both had a statistically significant impact on viscosity, according to a simple main effects analysis. A lower viscosity was found (p < .05) at 0.04 mg/mL (0.1 ± 0.0) after 10 min of incubation. A simple main effects analysis showed that papain concentrations and incubation time have a statistically significant effect on sperm motility (p = .0001). At 0.04 mg/mL papain, the sperm motility % was higher (p < .05) after 10 min (64.4 ± 4.8), 20 min (68.4 ± 6.2), and 30 min incubation (72.2 ± 6.6) compared to 0, 5 min (38.3 ± 4.1 and 51.6 ± 5.0, respectively). In conclusion, the fresh diluted camel semen had the lowest viscosity properties after 10 min of incubation with 0.04 mg/mL papain, without compromising sperm motility, viability, acrosome integrity and sperm morphology.

Comparing Nutritional Values and Bioactivity of Kefir from Different Types of Animal Milk.

The growing interest in fermented dairy products is due to their health-promoting properties. The use of milk kefir grains as a starter culture made it possible to obtain a product with a better nutritional and biological profile depending on the type of milk. Cow, buffalo, camel, donkey, goat, and sheep milk kefirs were prepared, and the changes in sugar, protein, and phenol content, fatty acid composition, including conjugated linoleic acids (CLAs), as well as antioxidant activity, determined by ABTS and FRAP assays, were evaluated and compared. The protein content of cow, buffalo, donkey, and sheep milk increased after 24 h of fermentation. The fatty acid profile showed a better concentration of saturated and unsaturated lipids in all fermented milks, except buffalo milk. The highest content of beneficial fatty acids, such as oleic, linoleic, and C18:2 conjugated linoleic acid, was found in the cow and sheep samples. All samples showed a better antioxidant capacity, goat milk having the highest value, with no correlation to the total phenolic content, which was highest in the buffalo sample (260.40 ± 5.50 µg GAE/mL). These findings suggested that microorganisms living symbiotically in kefir grains utilize nutrients from different types of milk with varying efficiency.

Microscopic and molecular investigation of vector borne haemoprotozoan diseases in dromedary camel of North Gujarat, India.

BACKGROUND OBJECTIVES: Vector-borne haemoprotozoan diseases comprise diverse group of single celled organism transmitted by haematophagus invertebrates. The current study was aimed at the identification of major haemoprotozoan (Babesia, Theileria and Trypanosoma) in dromedary camel of North Gujarat region in India using microscopy and Polymerase Chain Reaction (PCR). METHODS: A total of 234 blood samples were screened by the microscopic and molecular detection assays. Molecular prevalence studies of Theileria, Trypanosoma spp and Babesia was undertaken using 18s ribosomal DNA, RoTat 1.2 and SS rRNA gene respectively. The data relating to microscopic and molecular prevalence along with associated risk factors were analysed by statistical methods. RESULTS: The overall prevalence of hamoprotozoan disease based on microscopic and molecular investigation was 23.50%. The sensitivity and specificity (95% Confidence Interval) of PCR assay was 100% in comparison to microscopy (45.45 % sensitive and 100 % specific). The kappa coefficient between PCR and microscopy indicated good level of agreement with a value of 0.704 and SE of 0.159. INTERPRETATION CONCLUSION: Despite holding much significance to the animal sector, little work has been undertaken in regional parts of India regarding camel parasites. The present study offers first preliminary research data investigating haemoprotozoan disease using parasitological and molecular methods in camels in the region.

Characterizing Lomerizine metabolites in camel urine: High-resolution mass spectrometry method development and validation for enhanced doping control.

RATIONALE: Lomerizine (LMZ) is an antimigraine drug that works as a calcium channel blocker and has selective effects on the central nervous system. It is metabolized into trimetazidine (TMZ), which is a prohibited substance owing to its performance-enhancing effects in both human and animal sports. Effective doping control measures are imperative to distinguish the source of TMZ in samples to ensure integrity and fairness of the sport, therefore a comprehensive analysis of LMZ metabolites is essential to identify potential biomarkers in camel urine for effective doping control. METHODS: Camel urine samples were collected from four healthy animals following a single oral administration of LMZ at a dosage of 1 mg/kg body weight. In vitro studies were conducted using homogenized camel liver samples. Lomerizine and its metabolites were extracted using solid-phase extraction and analyzed with a Thermo Fisher Orbitrap Exploris liquid chromatography mass spectrometry system. The acquired data was processed with the Compound Discoverer software. RESULTS: The study conducted a comprehensive analysis of LMZ metabolites in camels and identified 10 phase I and one phase II metabolites. The primary pathway for the formation of phase I metabolites was de-alkylation, while phase II metabolite was formed through alkylation of the parent drug. The study provided valuable insights into the unique metabolic pathways of LMZ in camels under specific experimental conditions. CONCLUSION: The developed method enables the detection and characterization of LMZ and its metabolites in camels. The identified metabolites has the potential to act as marker metabolites for the distinctive detection of LMZ in camel urine to ensure efficient analytical strategies for routine doping control applications.

Serological and histolomorphological investigation of camel bulls testes (Camelus dromedaries) during the rutting and non-rutting seasons.

BACKGROUND: Camels are bred for their milk, meat, wool and hair, transportation, and their excrement as fuel. The seasonal reproduction of camel bull is accompanied by changes in sexual activity, the morphology, and function of the testes. This study aimed to evaluate the seasonal fluctuations in serum testosterone (T) levels as well as total antioxidant capacity (TAC) and malondialdehyde (MDA) levels in the testes of dromedary bulls (Camelus dromedarius) during the rutting and non-rutting seasons. Moreover, the impact of rutting season on the testicular size and histomorphology was also observed. Seventy mature dromedary bulls were divided into a rutting group (n = 35) and a non-rutting group (n = 35). From these bulls, blood samples and testes were collected during the rutting season (October to April) and non-rutting season (May to September) from a local slaughterhouse. RESULTS: All parameters changed significantly during rutting and non-rutting periods in camel bulls. The levels of TAC in testes, and serum T were significantly (P < 0.05) higher in the rutting group than in the non-rutting group. However, testicular MDA was significantly (P < 0.05) lower in the rutting group than in the non-rutting group. TAC was negatively correlated with MDA (r = -0.59, p < 0.01). Moreover, in the rutting group and the non-rutting group, T was positively correlated with levels of TAC (r = 0.66, p < 0.0003). Additionally, testicular size (length, breadth, and thickness) was significantly greater in camels during the rutting season than in camels during the non-rutting season. Moreover, the number and diameter of seminiferous tubules, and spermatogenesis increased during the rutting season, whereas, the collagen content and apoptosis increased during the non-rutting season. CONCLUSION: This study revealed that the rutting normal breeding season (NBS, rutting group) was associated with higher levels of total antioxidant capacity (TAC), T, and spermatogenic activity while the collagen content, concentrations of MDA (the oxidative stress factor) and apoptosis (an outcome of oxidative stress) were lower than those in the low breeding season (LBS, non-rutting group). In addition, the testicular size and seminiferous tubule diameter and number were higher during the NBS.

Trichuris Globulosa Von Linstow, 1901 from one-humped camel (Camelus dromedarius) in Egypt: prevalence, morphological and molecular study.

BACKGROUND: Trichuris spp. (whipworms) are soil-transmitted helminths distributed worldwide, parasitizing several mammalian hosts such as ruminants, primates, and rodents. Trichuris spp. is one of the most common intestinal parasites affecting both humans and animals, and it can spread directly through the fecal-oral route, resulting in severe illness and financial loss. So, this work aims to detect the frequency of Trichuris spp. in camels in Beheira Governorate, Egypt, and to identify Trichuris spp. through morphometrical studies, molecular analysis, and phylogenetic analysis. RESULTS: A total of 35 dromedaries out of 127 investigated had Trichuris spp. infection, meaning that the overall prevalence was 27.56%. The age of the camel affected the infection rate, older animals (> 5 years) having a higher prevalence of infection (24%) than animals of ages (< 3 years) (20%) than animals of ages (3-5 years) (19.14%). According to season: Trichuris spp. showed a unique pattern in camels in different seasons: summer (31.25%) > autumn (28.13%) > spring (25.8%) > winter (25%) indicating year-round infection. T. globulosa was identified morphometrically from camels in Beheira Governorate, Egypt. The BLAST analysis revealed the presence of T. globulosa isolate from camels using the Genbank database depending on nuclear small subunit ribosomal RNA (18s) and cytochrome b (Cytb) genes. CONCLUSION: A high prevalence of T. globulosa was found in camels in Beheira Governorate, Egypt. This is the first report to confirm the identification of T. globulosa from camel based on morphometrical studies and molecular and phylogenetic analysis in Egypt. More thorough studies on the incidence, molecular, and genetic analysis of Trichuris spp. in Egypt are required in addition to camel control programs.