Molecular epidemiology and associated risk factors of Anaplasma marginale in camels and possible co-infections.

Anaplasma spp. is an important pathogen that affects a wide range of animals, including camels. The current study aimed to assess the prevalence of six Anaplasma spp. in 400 camels from Ismailia, Suez, and Sharkia governorates in northern Egypt, as well as their associated risk factors and possible coinfections. Blood and fecal samples were examined using bacterial culture, the vitek2 system, and PCR. Genetic divergence among Anaplasma marginale (A. marginale) isolates was characterized using the msp4 gene. The overall prevalence of A. marginale was 19.5%. Sequencing analysis confirmed the PCR results, and a single A. marginale genotype was recognized by msp4 sequencing. The phylogenetic tree indicated that the study A. marginale isolates clustered together and were close to Egyptian A. marginale identified from buffalo (OP142725 and OP142726). Age, sex, housing type, tick infestation, body conditions, and tick control factors were significantly associated with camel anaplasmosis using a logistic regression model (odds ratio >1, P < 0.05). Multivariate logistic regression analysis revealed that the infection was 2.03, 1.9, 2.6, 1.9, and 1.8 times higher in females, semi-enclosed housing, ages >5 years, tick infestation, and emaciated camels. The risk of infection due to a tick control factor increased by 4.4 and 2.6 times when no control was applied or with irregular control, respectively. This is the first molecular report of A. marginale infection in camels in Ismailia, Suez, and Sharkia in northern Egypt, indicating a moderate prevalence of A. marginale and the involvement of multiple bacterial infections, mainly Escherichia coli and Salmonella spp. Thus, it is necessary to develop effective management and control for camel anaplasmosis.

Investigating Crimean-Congo haemorrhagic fever virus seropositivity in camels and human behavioural risks in an abattoir in Nigeria.

Crimean-Congo haemorrhagic fever virus (CCHFV) is an emerging viral pathogen with pandemic potential that is often misdiagnosed. Case fatality in low-resource settings could be up to 40% due to close contact between animals and humans. A two-year cross-sectional study was conducted in Fagge abattoir, Kano State, Nigeria, to estimate the seropositivity of CCHFV in camels using a commercial multi-species competitive enzyme-linked immunosorbent assay (ELISA). A closed-ended questionnaire was administered to the abattoir workers to assess their awareness, mitigation, and behavioural practices associated with CCHF. Of the 184 camels tested, 179 (97%) were seropositive for CCHFV (95% confidence interval (CI): 93.77, 99.11). The median (interquartile range (IQR)) age of respondents was 41 (35-52), with 62% having no education. Respondents had little knowledge about CCHFV and the concept of zoonotic disease. In this study, the high estimated prevalence of antibodies to CCHFV in camels highlights the heightened risk of transmission of CCHFV in Nigeria. Similarly, a concerning lack of knowledge and inadequate preventive practices, alongside a prevalence of high-risk behaviours associated with CCHF among abattoir workers, were noted in this study. Thus, there is an urgent need for comprehensive public health education and collaborative One Health strategies to avert the threats of spillover events.

Grazing camels under semi-extensive production system: selectivity, feed intake capacity, digestion and energy expenditure.

BACKGROUND: It was proposed that camels are more effective than other livestock species in selecting plants for their nutritional value. They may self-regulate their voluntary feed intake to satisfy their nutritional needs. This study was designed to investigate camels' feeding selectivity and ability to cover nutritional requirements when grazing alfalfa (high in protein) and/ or barley (high in energy) in a desert climate. METHODS: Eighteen lactating camels were equally divided into three feeding treatments. They grazed daily on alfalfa, barley, or a mixed pasture of both, for two periods of one month each. The concentrate supplement was individually administered at 40 g/kg BW0.75, divided into two equal parts, in the morning and in afternoon. Total energy expenditure (EE) was estimated by heart rate (HR) monitors for 48 h after being calibrated by oxygen consumption using an upgraded face mask open-circuit respiratory system. RESULTS: During the first period, camels had a greater forage intake and digestibility when they grazed barley rather than alfalfa, while those grazing mixed pasture performed intermediately. In the second period, camels had a similar forage intake and digestibility among treatments due to a decline in barley intake and digestibility compared to the first period, which was expected since the preferred plant part gradually shifted from barley grains to predominantly straw as a function of time. Similar HR and EE were found across periods and treatments. As a result of greater gross and digestible energy intake in period 1, a better energy balance in period 1 was observed compared to period 2. Camels better utilize barley than alfalfa. Grazing on barley had a higher energy balance than grazing alfalfa alone or in combination with barley. However, camels grazing barley produced lower milk yield and energy than those grazing alfalfa alone or in combination with barley, with no interaction detected between period and treatment. CONCLUSIONS: Lactating camels are able to self-regulate their voluntary intake to cover their energy requirements when they are grazing barley and/or alfalfa supplemented with a concentrate supplement at 40 g/kg BW0.75. Grazing barley is better utilized by camels than alfalfa. The chemical and physical properties of plant species play an important role in the selectivity of foraging camels. It also impacts their intake and digestibility, which is negatively associated with the proportion of cell wall content consumed.

Developmental characteristics of aggregated lymphoid nodules area in the abomasum of fetal Bactrian camels (Camelus bactrianus).

BACKGROUND: Bactrian camel is one of the important economic animals in northwest China. They live in arid desert, and their gestation period is about 13 months, which is longer than other ruminants (such as cattle and sheep). The harsh living conditions have made its unique histological characteristics a research focus. Aggregated lymphoid nodules area (ALNA) in the abomasum of Bactrian camels, as one of the most important sites for the induction of the immune response, provide a comprehensive and effective protective role for the organism, and their lack of information will affect the feeding management, reproduction and epidemic prevention of Bactrian camels. In this study, the histological characteristics of the fetal ALNA in the abomasum of Bactrian camels at different developmental gestation have been described by using light microscopy and histology . RESULTS: The ALNA in the abomasum of the Chinese Alashan Bactrian camel is a special immune structure that was first discovered and reported by Wen-hui Wang. To further establish the developmental characteristics of this special structure in the embryonic stage, the abomasum ALNA of 8 fetuses of Alashan Bactrian camels with different gestational ages (5~13 months) were observed and studied by anatomy and histology. The results showed that the aggregation of reticular epithelial cells (RECs) surrounded by a very small number of lymphoid cells was detected for the first time in the abomasum of fetal camel at 5 months gestation, which was presumed to be primitive ALNA. At 7 months gestation, the reticular mucosal folds region (RMFR) appeared, but the longitudinal mucosal folds region (LMFR) was not significant, and histological observations showed that there were diffusely distributed lymphocytes around the RECs. At 10months gestation, RMFR and LMFR were clearly visible, lymphoid follicles appeared in histological observation, lymphocytes proliferated vigorously. By 13 months, the volume of lymphoid follicles increased, forming the subepithelial dome (SED), and there was a primitive interfollicular area between the lymphoid follicles, which contained high endothelial vein (HEV), but no germinal center (GC) was found. In summary, ALNA of Bactrian camels is not fully mature before birth. CONCLUSIONS: Generally, the small intestine PPs of ruminants (such as cattle and sheep) is already mature before birth, while the ALNA in the abomasum of Bactrian camels is not yet mature in the fetal period. During the development of ALNA in Bactrian camel, the development of lymphoid follicles extends from submucosa to Lamina propria. Interestingly, the deformation of FAE changes with age from simple columnar epithelium at the beginning of pregnancy to Simple cuboidal epithelium, which is opposite to the FAE deformation characteristics of PPs in the small intestine of fetal cattle and sheep. These results are the basis of further research on the specificity of ALNA in the abomasum of Bactrian camels.

Camel filariasis (Dipetalonema evansi) and its association with clinical balanoposthitis with reference to prominent changes in clinical findings, serum testosterone, semen analysis, and testicular histopathology.

BACKGROUND: Camel filariasis induced variable clinical syndromes characterized by fever, lethargy, localized dermal lesions, loss of condition, and testicular and scrotal swelling. The objective of the present work focused on clarifying the diagnostic importance of clinical findings, serum testosterone, and semen analysis as well as blood smear and testicular histopathology as a differential tool between only balanoposthitis without filariasis male camels group (OnlyBpgr) and balanoposthitis-filariasis infected male camels group (BpFlgr). The study also monitored the associations between the severity of ticks' infestations in investigated male camels and the occurrence of balanoposthitis only or balanoposthitis with filariasis. RESULTS AND CONCLUSIONS: The study reported significant correlation between serum testosterone, serum cortisol, and sperm vitality and abnormalities percentages. The study included male camels (n = 250) classified into three groups: healthy control group (Contgr; n = 30), OnlyBpgr (n = 210), and BpFlgr (n = 10). These male camels were clinically and laboratory examined, and skin scraping tests and testicular histopathology were conducted. The study confirmed the association of the changes in clinical findings, whole blood picture, serum testosterone, serum cortisol, and semen analysis, with OnlyBpgr and BpFlgr. These changes were more prominent in BpFlgr than in OnlyBpgr. Skin scraping test results revealed a higher severity of live ticks' infestation in BpFlgr than in OnlyBpgr because, unlike OnlyBpgr, all camels in BpFlgr (n = 10) were suffering from live ticks' infestation. It also concluded the higher efficacy of histopathology of testicular tissues in male camels as a diagnostic tool for adult filaria in balanoposthitis-affected male camels than blood smear because all cases of camel filariasis in the current work were negative for microfilaria on microscopic examination of diurnal blood smear as well as testicular histopathology revealed detection of adult filaria in all camel filariasis associated with balanoposthitis. Strong correlation relationships were demonstrated between serum testosterone, serum cortisol, and semen analysis results. Positive correlations were reported between serum testosterone levels and sperm vitality percentages. However, negative correlations were stated between serum testosterone and each of serum cortisol and sperm abnormalities either in Contgr, OnlyBpgr, or BpFlgr.

Effects of long-term dehydration and quick rehydration on the camel kidney: pathological changes and modulation of the expression of solute carrier proteins and aquaporins.

BACKGROUND: Recurrent dehydration causes chronic kidney disease in humans and animal models. The dromedary camel kidney has remarkable capacity to preserve water and solute during long-term dehydration. In this study, we investigated the effects of dehydration and subsequent rehydration in the camel's kidney histology/ultrastructure and changes in aquaporin/solute carrier proteins along with gene expression. RESULTS: In light microscopy, dehydration induced few degenerative and necrotic changes in cells of the cortical tubules with unapparent or little effect on medullary cells. The ultrastructural changes encountered in the cortex were infrequent during dehydration and included nuclear chromatin condensation, cytoplasmic vacuolization, mitochondrial swelling, endoplasmic reticulum/ lysosomal degeneration and sometimes cell death. Some mRNA gene expressions involved in cell stability were upregulated by dehydration. Lesions in endothelial capillaries, glomerular membranes and podocyte tertiary processes in dehydrated camels indicated disruption of glomerular filtration barrier which were mostly corrected by rehydration. The changes in proximal tubules brush borders after dehydration, were accompanied by down regulation of ATP1A1 mRNA involved in Na + /K + pump that were corrected by rehydration. The increased serum Na, osmolality and vasopressin were paralleled by modulation in expression level for corresponding SLC genes with net Na retention in cortex which were corrected by rehydration. Medullary collecting ducts and interstitial connective tissue were mostly unaffected during dehydration. CKD, a chronic nephropathy induced by recurrent dehydration in human and animal models and characterized by interstitial fibrosis and glomerular sclerosis, were not observed in the dehydrated/rehydrated camel kidneys. The initiating factors, endogenous fructose, AVP/AVPR2 and uric acid levels were not much affected. TGF-ß1 protein and TGF-ß1gene expression showed no changes by dehydration in cortex/medulla to mediate fibrosis. KCNN4 gene expression level was hardly detected in the dehydrated camel's kidney; to encode for Ca + + -gated KCa3.1 channel for Ca + + influx to instigate TGF-ß1. Modulation of AQP 1, 2, 3, 4, 9 and SLC protein and/or mRNAs expression levels during dehydration/rehydration was reported. CONCLUSIONS: Long-term dehydration induces reversible or irreversible ultrastructural changes in kidney cortex with minor effects in medulla. Modulation of AQP channels, SLC and their mRNAs expression levels during dehydration/rehydration have a role in water conservation. Cortex and medulla respond differently to dehydration/rehydration.

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.

Spatiotemporal convolutional long short-term memory for regional streamflow predictions.

Rainfall-runoff (RR) modelling is a challenging task in hydrology, especially at the regional scale. This work presents an approach to simultaneously predict daily streamflow in 86 catchments across the US using a sequential CNN-LSTM deep learning architecture. The model effectively incorporates both spatial and temporal information, leveraging the CNN to encode spatial patterns and the LSTM to learn their temporal relations. For training, a year-long spatially distributed input with precipitation, maximum temperature, and minimum temperature for each day was used to predict one-day streamflow. The trained CNN-LSTM model was further fine-tuned for three local sub-clusters of the 86 stations, assessing the significance of fine-tuning in model performance. The CNN-LSTM model, post fine-tuning, exhibited strong predictive capabilities with a median Nash-Sutcliffe efficiency (NSE) of 0.62 over the test period. Remarkably, 65% of the 86 stations achieved NSE values greater than 0.6. The performance of the model was also compared to different deep learning models trained using a similar setup (CNN, LSTM, ANN). An LSTM model was also developed and trained individually to predict for each of the stations using local data. The CNN-LSTM model outperformed all the models which was trained regionally, and achieved a comparable performance to the local LSTM model. Fine-tuning improved the performance of all models during the test period. The results highlight the potential of the CNN-LSTM approach for regional RR modelling by effectively capturing complex spatiotemporal patterns inherent in the RR process.

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.

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.

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.

Crimean-Congo Hemorrhagic Fever Virus Seropositivity among Dromedary Camels, Algeria, 2020-2021.

Serosurvey results for Crimean-Congo hemorrhagic fever virus antibodies in dromedary camels in Algeria indicate that the pathogen is circulating endemically in desertic areas, despite the hostile environment. Thus, dromedaries are suitable sentinels for detecting human risk for Crimean-Congo hemorrhagic fever in desertic areas.

MERS-CoV‒Specific T-Cell Responses in Camels after Single MVA-MERS-S Vaccination.

We developed an ELISPOT assay for evaluating Middle East respiratory syndrome coronavirus (MERS-CoV)‒specific T-cell responses in dromedary camels. After single modified vaccinia virus Ankara-MERS-S vaccination, seropositive camels showed increased levels of MERS-CoV‒specific T cells and antibodies, indicating suitability of camel vaccinations in disease-endemic areas as a promising approach to control infection.

Large-scale application of palaeoproteomics (Zooarchaeology by Mass Spectrometry; ZooMS) in two Palaeolithic faunal assemblages from China.

The application of Zooarchaeology by Mass Spectrometry (ZooMS) on Pleistocene sites in Europe and northern Asia has resulted in the discovery of important new hominin fossils and has expanded the range of identified fauna. However, no systematic, large-scale application of ZooMS on Palaeolithic sites in East Asia has been attempted thus far. Here, we analyse 866 morphologically non-diagnostic bones from Jinsitai Cave in northeast China and Yumidong Cave in South China, from archaeological horizons dating to 150-10 ka BP. Bones from both sites revealed a high degree of collagen preservation and potentially time-related deamidation patterns, despite being located in very distinct environmental settings. At Jinsitai, we identified 31 camel bones, five of which were radiocarbon dated to 37-20 ka BP. All dated specimens correspond to colder periods of Marine Isotope Stages 3 and 2. We regard the presence of camels at Jinsitai as evidence of wild camels being a megafauna taxon targeted, most likely by early modern humans, during their expansion across northeast Asia. This large-scale application of ZooMS in China highlights the potential of the method for furthering our knowledge of the palaeoanthropological and zooarchaeological records of East Asia.

Molecular prevalence, associated risk factors and genetic characterization of Trypanosoma evansi in camels.

Surra is a major infectious disease of camels being caused by Trypanosoma evansi (T. evansi) in developing countries, including Egypt. However, the identification of changes in the T. evansi prevalence in Egypt is important. In this study, the prevalence of T. evansi and its associated risk factors as well as the genetic characterization of the parasite were estimated. Blood samples were collected from 163 camels from two governorates in Lower Egypt. PCR targeting RoTat 1.2VSG was used for the detection of T. evansi and internal transcribed spacer 1 (ITS-1) was used for sequencing analysis and genetic characterization. Overall prevalence was 19.6% using RoTat 1.2VSG. The risk of the infection in females was 4 times higher than in males (P = 0.0004, OR = 4; 95% CI = 0.79-8.96) and in camels with a history of clinical signs it was 2.3 times higher than camels without clinical signs (P = 0.04, OR = 2.3, 95% CI = 1.035-5.15). Analysis of the ITS-1 sequences of four T. evansi isolates showed little heterogeneity compared to similar sequences in the database. Sequence and phylogenetic analysis, based on the ITS-1 region, confirmed the presence of two distinct genotypes of T. evansi in Egyptian camels with more than 99% similarity with T. evansi isolates from different countries across the ITS-1 region and were closely related to Filipino and Chinese isolates. The results of the study can be used for the observation and prevention of disease and updating the epidemiological data.

Detection of viral antibodies in camel sera using magnetic particle spectroscopy.

Pandemics like SARS-Cov-2 very frequently have their origin in different animals and in particular herds of camels could be a source of zoonotic diseases. This study took advantage on a highly sensitive and adaptable method for the fast and reliable detection of viral antibodies in camels using low-cost equipment. Magnetic nanoparticles (MNP) have high variability in their functionalization with different peptides and proteins. We confirm that 3-aminopropyl triethoxysilane (APTES)-coated MNP could be functionalized with viral proteins. The protein loading could be confirmed by simple loading controls using FACS-analysis (p < 0.05). Complementary combination of antigen and antibody yields in a significant signal increase could be proven by both FACS and COMPASS. However, COMPASS needs only a few seconds for the measurement. In COMPASS, the phase φn on selected critical point of the fifth higher harmonic (n = 5th). Here, positive sera display highly significant signal increase over the control or negative sera. Furthermore, a clear distinction could be made in antibody detection as an immune response to closely related viruses (SARS-CoV2 and MERS). Using modified MNPs along with COMPASS offers a fast and reliable method that is less cost intensive than current technologies and offers the possibility to be quickly adapted in case of new occurring viral infections. KEY POINTS: • COMPASS (critical offset magnetic particle spectroscopy) allows the fast detection of antibodies. • Magnetic nanoparticles can be adapted by exchange of the linked bait molecule. • Antibodies could be detected in camel sera without washing steps within seconds.

Antibody preparation and age-dependent distribution of TLR8 in Bactrian camel spleens.

BACKGROUND: Toll-like receptor 8 (TLR8) can recognize specific pathogen-associated molecular patterns and exert multiple immunological functions through activation of signaling cascades. However, the precise distribution and age-related alterations of TLR8 in the spleens of Bactrian camels have not yet been investigated. This study aimed to prepare a rabbit anti-Bactrian camel TLR8 polyclonal antibody and elucidate the distribution of TLR8 in the spleens of Bactrian camels at different age groups. The methodology involved the construction of the pET-28a-TLR8 recombinant plasmid, followed by the expression of TLR8 recombinant protein via prokaryotic expression. Subsequently, rabbits were immunized with the purified protein to prepare the TLR8 polyclonal antibody. Finally, twelve Alashan Bactrian camels were categorized into four groups: young (1-2 years), pubertal (3-5 years), middle-aged (6-16 years) and old (17-20 years). These camels received intravenous sodium pentobarbital (20 mg/kg) anesthesia and were exsanguinated to collect spleen samples. Immunohistochemical techniques were employed to observe and analyze the distribution patterns and age-related changes of TLR8 in the spleen. RESULTS: The results showed that the TLR8 recombinant protein was expressed in the form of inclusion body with a molecular weight of 52 kDa, and the optimal induction condition involved 0.3 mmol/L IPTG induction for 8 h. The prepared antibody yielded a titer of 1:32 000, and the antibody demonstrated specific binding to TLR8 recombinant protein. TLR8 positive cells exhibited a consistent distribution pattern in the spleen across different age groups of Bactrian camels, primarily scattered within the periarterial lymphatic sheath of the white pulp, marginal zone, and red pulp. The predominant cell type expressing TLR8 was macrophages, with expression also observed in neutrophils and dendritic cells. Statistical analysis revealed that there were significant differences in the distribution density of TLR8 positive cells among different spleen regions at the same age, with the red pulp, marginal zone, and white pulp showing a descending order (P<0.05). Age-related changes indicated that the distribution density in the marginal zone and red pulp exhibited a similar trend of initially increasing and subsequently decreasing from young to old camels. As camels age, there was a significant decrease in the distribution density across all spleen regions (P<0.05). CONCLUSIONS: The results confirmed that this study successfully prepared a rabbit anti-Bactrian camel TLR8 polyclonal antibody with good specificity. TLR8 positive cells were predominantly located in the red pulp and marginal zone of the spleen, signifying their pivotal role in the innate immune response of the spleen. Aging was found to significantly reduce the density of TLR8 positive cells, while leaving their scattered distribution characteristics unaffected. These findings provide valuable support for further investigations into the immunomorphology and immunosenescence of the spleen in Bactrian camels.

Effect of concentrates level on digestibility, ruminal fermentation, and bacterial community in growing camels.

Understanding the rumen microbiota of camels under different feeding conditions is necessary to optimize rumen fermentation and productivity. This study aims to investigate the effects of different concentrate supplement levels on digestion, rumen fermentation and bacteria in growing camels. Fifteen growing camels were divided into three groups and were fed alfalfa hay in addition to one of the three concentrate supplement levels based on body weight (BW): low (0.7%), medium (1%), and high (1.3%). Increasing the concentrate supplement level in the diet increased total dry matter intake but had no effect on nutrients digestibility, except for crude protein digestibility, which was enhanced with the high concentrate level. Growing camels at low-level had considerably higher rumen pH than those fed medium or high levels. Increasing the supplement level also increased rumen propionic acid but decreased acetic acid concentration. Principal coordinate analysis showed that concentrate levels clearly separated the ruminal bacterial communities where Bacteroidetes and Firmicutes were the dominant phyla and Prevotella, Ruminococcus, Butyrivibrio, RC9_gut_group, and Fibrobacteres were the dominant bacterial genera. This study expands our knowledge regarding the rumen microbiota of growing camels under different concentrate levels and reveals that medium concentrate levels could be appropriate for growing camels.

Comparative digestibility and rumen fermentation of camels and sheep fed different forage sources.

The present study evaluated the effects of forage sources on dry matter (DM) intake, digestibility, and fermentation parameters in camels vs. sheep. The study was arranged as a 2 × 3 factorial experiment in a completely randomized design by using two animal species (three ruminally cannulated female camels and three male sheep) and three forage sources. The forages were (1) alfalfa hay; 164 and 479 g/kg DM for crude protein (CP) and neutral detergent fiber (NDF), respectively, (2) berseem hay; 121 and 513 g/kg DM for CP and NDF, respectively, and (3) wheat straw (27.5 and 723 g/kg DM for CP and NDF, respectively). Higher DM intake [g/kg body weight (BW)] was noted in sheep compared to camel when alfalfa hay (p < 0.05) and berseem hay (p < 0.05) were fed but was similar between both species when they were fed wheat straw. Forage type rather than animal species had more effect on metabolic intake. Lower digestibility was noted in sheep with wheat straw (p < 0.05). Similar in situ degradability values for crude protein and DM were noted between camels and sheep. In situ degradability of NDF was higher (p < 0.05) in camel than sheep. Greater ruminal pH (p < 0.05) was noted in sheep vs. camels when berseem hay and wheat straw were fed. Lower ruminal passage rate (p < 0.05) was noted in camels and higher total mean retention time. It was concluded that sheep and camels had similar digestion capacities when fed berseem hay and alfalfa hay, however, camels are more efficient than sheep when fed wheat straw.

Molecular screening of Amblyomma species (Acari: Ixodidae) imported from African countries to Egypt, with the first report of Amblyomma latum from the ball python, Python regius (Squamata: Pythonidae).

Amblyomma species are non-endemic ticks in Egypt, which have been recorded from imported animals. This study was carried out in 2022 to monitor Amblyomma spp. from dromedary camels, cattle, and snakes in Egypt. During this study, 400 camels, 200 cattle, and two snakes (Pythonidae) were inspected for tick infestation. Collected specimens were identified based on morphological characters and confirmed by phylogenetic analysis of the 12S rRNA gene. Camels were infested by adult specimens of Amblyomma variegatum and Amblyomma lepidum, but no Amblyomma spp. were collected from cattle. Amblyomma variegatum showed high genetic similarity to other A. variegatum from Guinea-Bissau and São Tomé (> 99.99%), and A. lepidum showed high genetic similarity to other A. lepidum from Israel and Sudan (99.99%). Amblyomma latum is recorded in Egypt from the ball python snake for the first time and showed high genetic similarity with South African A. latum (99.87%).