Antibacterial, Anti-Biofilm, and Anti-Inflammatory Properties of Gelatin-Chitosan-Moringa-Biopolymer-Based Wound Dressings towards Staphylococcus aureus and Escherichia coli.

In contemporary times, the sustained aspiration of bioengineering and biomedical applications is the progressive advancement of materials characterized by biocompatibility and biodegradability. The investigation of the potential applications of polymers as natural and non-hazardous materials has placed significant emphasis on their physicochemical properties. Thus, this study was designed to investigate the potential of gelatin-chitosan-moringa leaf extract (G-CH-M) as a novel biomaterial for biomedical applications. The wound-dressing G-CH-M biopolymer was synthesized and characterized. The blood haemolysis, anti-inflammatory, antioxidant, and antibacterial activities of the biopolymer were investigated against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacterial isolates. Our results showed that S. aureus swarming motility was drastically affected. However, the biopolymer had no significant effect on the swarming motility of E. coli. In addition, the biopolymer showed high antibacterial capacities, especially against S. aureus. Plasmid DNA was observed to be effectively protected from oxidative stresses by the biopolymer. Furthermore, the biopolymer exhibited greatly suppressed haemolysis (lower than 2%), notwithstanding the elevated concentration of 50 mg/mL. These results indicated that this novel biopolymer formulation could be further developed for wound care and contamination prevention.

Inhibitory potential of natural plant extracts against Escherichia coli strain isolated from diarrheic camel calves.

Background: Camel calf's diarrhea is considered the chief economic loss in the camelid population. There is currently no vaccine licensed to prevent colibacillosis in camel calves. The new era of bacterial antibiotic resistance explains the treatment failure and the high mortality and morbidity associated with the disease. Current protective treatments have thus far limited efficacy and need to be replaced. Due to their antimicrobial properties and safety, natural products are recently finding a capital role in infection management. Aims: The current study explores Escherichia coli F17 susceptibility as a clinical strain isolated from diarrheic camel calves to a wide panel of natural products. Methods: Agar diffusion method, integrity of cell membrane, hydrophobicity of bacterial surface, biofilm assays, and motility were used to evaluate the antibacterial activity of Coffea, Retama raetam, Moringa oleifera, Juniperus phoenicea, Uritica dioica, Camellia sinensis, Lavandula angustifolia, and Cuminum cyminum extracts against isolated bacteria. Results: Interestingly, all eight tested extracts have the damaging ability of E. coli F17's cell membrane and cause the nucleic acid release after 12 hours. Escherichia coli F17 strain has the surface of hydrophobicity which changed after contact with extracts of the plant. Moreover, the motility of the studied bacteria changed after exposure to all plant extracts. Conclusion: This study demonstrated that all extracts, exempt U. dioica, can remove up to 50% biofilm of E. coli biomass as compared with the control. Natural extracts can be used as potential antimicrobial agents to mitigate diarrhea in camel calves.

Usage of the XGBoost and MARS algorithms for predicting body weight in Kajli sheep breed.

This study aimed to utilize the XGBoost and MARS algorithms to predict present weight from body measurements. The algorithms have the potential to model nonlinear relationships between body measurements and weight, and this study attempted to find a model that provided the most accurate predictions of present weight. The current study was conducted with 152 animals in order to achieve a certain goal. To compare the model performances, goodness-of-fit criteria such as R2, r, RMSE, CV, SDratio, PI, MAPE, AIC were used. According to the results of this study, the XGBoost algorithm was the most reliable model for predicting present weight from body measurement. Even if the XGBoost algorithm was the most accurate model, the MARS algorithm was the reliable model for the same aim. In addition, it is hoped that the results of this study will help researchers and breeders better understand the relationship between body measurements and weight and ultimately be able to help individuals better manage their weight. As a conclusion, in the current study, the XGBoost algorithm is an effective, efficient, and reliable tool for accurately estimating present weight from body measurements. This makes it an invaluable tool in rural areas, where traditional weighing scales may not be available or reliable.

European Regulations on Camel Germplasm Movement within the European Union: A Current Framework Based on Safety.

With the aim of developing livestock breeding, the Delegated Regulation (EU) 2020/686 (hereafter referred to as Reg. 686) has taken steps to define traceability and animal health for the movement of germ material within the European Union (EU), including that of camelid species. Despite the economic importance of the camel market and the efforts of the EU to regulate their movements, there are considerable difficulties in the collection of semen and its freezing, limiting the use of artificial insemination in this species. If, on the one hand, there is little diffusion of the camel breeding and, consequently, limited diffusion of animals and germplasm, there will probably be a significant increase over the years. To avoid the spread of emerging diseases-or even those no longer present in Europe-the entry of genetic material from non-EU countries must be strictly monitored. Camels are rarely clinically compliant, but can transfer even fatal diseases to domestic ungulate farms in the EU. Based on these considerations, we conducted a narrative review of the European regulations on this issue, focusing on aspects related to their application in camels.

Detection of Trypanosoma Infection in Dromedary Camels by Using Different Diagnostic Techniques in Northern Oman.

Camel trypanosomoses is considered a devastating disease with severe health consequences that can be caused by different hemoprotozoan parasites. Camel samples (388) from the five regions in Northern Oman were assessed using a thin blood film. In addition, 95 seropositive samples were analyzed using various primers of mechanically transmitted trypanosomes. Out of the 388 blood smears examined, 0.8% (CI 95%, 2/388) were found to be positive for Trypanosoma sp. using a microscope. The parasitologically positive cases were detected in samples from females. The overall molecular prevalences were as follows: TBR was 78/95, 77% (CI 73.1-89.2%); ITS was 30/95, 31.6% (CI 73.1-89.2%); and T. evansi type A (RoTat 1.2) was 8/95, 8.4% (CI 4.0-16.0%). There were two species of trypanosomes that were observed in the camels.

Identification of Candidate Genes for Pigmentation in Camels Using Genotyping-by-Sequencing.

The coat color of dromedary is usually uniform and varies from black to white, although dark- to light-brown colors are the most common phenotypes. This project was designed to gain knowledge on novel color-related variants using genotyping-by-sequencing (GBS). The association between the SNPs and coat color was tested using MLM (mixed linear models) with kinship matrix. Three GWAS models including white color vs. non-white color, black vs. non-black color, and light-brown vs. dark-brown color were performed. There were no distinct genetic clusters detected based on the color phenotypes. However, admixture occurred among all individuals of the four different coat color groups. We identified nine significant SNPs associated with white color after Bonferroni correction, located close to ANKRD26, GNB1, TSPYL4, TEKT5, DEXI, CIITA, TVP23B, CLEC16A, TMPRSS13, FXYD6, MPZL3, ANKRD26, HFM1, CDC7, TGFBR3, and HACE1 genes in neighboring flanking regions. The 13 significant SNPs associated with black color and the candidate genes were: CAPN7, CHRM4, CIITA, CLEC16A, COL4A4, COL6A6, CREB3L1, DEXI, DGKZ, DGKZ, EAF1, HDLBP, INPP5F, MCMBP, MDK, SEC23IP, SNAI1, TBX15, TEKT5, TMEM189, trpS, TSPYL4, TVP23B, and UBE2V1. The SNAI1 gene interacted with MCIR, ASIP and KIT genes. These genes play a key role in the melanin biosynthetic and pigmentation biological process and melanogenesis biological pathway. Further research using a larger sample size and pedigree data will allow confirmation of associated SNPs and the identified candidate genes.

Prevalence of Aflatoxins in Selected Dry Fruits, Impact of Storage Conditions on Contamination Levels and Associated Health Risks on Pakistani Consumers.

Dry fruits and nuts are nutritious foods with several health-promoting properties. However, they are prone to contamination with aflatoxins at all stages of production and storage. The present study aimed to determine the natural occurrence of aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), and total aflatoxins (AFT) in dates, pistachios, and walnuts collected from four districts of South Punjab (Pakistan), and to assess the associated health risks as estimated by dietary exposure and the Margin of Exposure (MoE) determinations. The contents of AFB1 and AFT in these food products were monitored during storage under three different conditions (open-air, hermetically closed jars, and refrigeration at 4 °C) to determine the most efficient conditions in preventing aflatoxin accumulation. HPLC-fluorescence analysis of 60 samples of these products for aflatoxin contamination showed that 52 (86.7%) samples were contaminated at different levels, with a maximum of 24.2 ng/g. The overall (all samples) mean concentrations of AFB1, AFB2, AFG1, AFG2, and AFT were 3.39 ± 2.96, 1.39 ± 1.68, 1.63 ± 1.48. 1.12 ± 1.23, and 7.54 ± 6.68, respectively. The Estimated Daily Intake (EDI) and MoE of aflatoxins through the consumption of the products ranged from 0.06 ng/kg bw/day to 2.0 ng/kg bw/day and from 84.84 to 2857.13, respectively, indicating that consumers are at high health risk. Significant differences were recorded between aflatoxin levels in the samples stored under different storage conditions, with storage under refrigeration (4 °C) being the most effective in controlling aflatoxin accumulation, although storage in closed jars was also efficient and offers a more flexible alternative to retailers. The findings of the study urge official authorities of Pakistan to implement appropriate regulatory and control measures and surveillance program to alleviate the potential public health risks associated with the consumption of dry fruits and nuts in the scope of their increased consumption.

Impact of various watering regimes on physiological and hematological parameters in intensively kept Marecha (Camelus dromedarius) she-camels in summer season.

Background: Camel is vital to daily life as a source of food and means of transportation, and just as importantly, its milk and urine have been used as medicine for diverse ailments since ancient times for people living in the desert region. Aim: The current study was planned to investigate the effect of different watering regimes on physiological and hematological parameters of lactating Marecha camels during the summer season. Methods: For this purpose, 12 lactating she-camels of almost similar age (8-13 years) and parity (2-5) were selected in a completely randomized design with four animals per treatment at the Camel Breeding and Research Station Rakh Mahni District Bhakkar. The trial was completed in 60 days, while 7 days were given as an acclimatization period for adaptation. The treatments were as follows: Group-1 (G-1) considered as the control had access to water once in a day; Group-2 (G-2) had access to water once in 4 days, and Group-3 (G-3) had access to water once in 6 days. Animals were housed individually for feeding and watering in an intensive management system. The same feed consisting of green fodder (Lucerne) and gram straw (Cicer arientinum) was offered to all animals. The average ambient temperature and relative humidity were 39°C-41°C and 55%-63%, respectively. Result: The mean values (p < 0.001) of water intake were found to be 82.94 ± 1.34 l in G-3, which was higher than G-1 and G-2. Blood analysis showed that packed cell volume, white blood cells, and hemoglobin were affected significantly (p < 0.001, p < 0.01, and p < 0.05, respectively). Conclusion: No serious changes in complete blood count and physiological parameters were reported in this study. So, this shows that the camel is unique in its physiological adaptation in hot and hostile environments.

Effect of Different Watering Regimes in Summer Season on Water Intake, Feed Intake, and Milk Production of Marecha She-camel (Camelus dromedarius).

Twelve lactating healthy Marecha she-camels in the early lactation stage during the summer at Camel Breeding and Research Station Rakh-Mahni (Pakistan) were included. All animals were fed with Medicago sativa and Cicer arientinum ad libitum and divided into three groups in relation to the access to water, after a period of seven days of adaptation to experimental conditions. Group 1 (G1) was considered as control having access to water once every day; Group 2 (G2) had access once every 4 days, while Group 3 (G3) had access once every 6 days. The duration of the study was 60 days with an adaptation period to experimental conditions of 7 days before the onset of the study. Dry matter intake (DMI) was calculated on a dry matter basis. On average the ambient temperature and relative humidity during the trial were 39-41 °C and 55-63%, respectively. The DMI, water intake, milk production, and body weight changes were affected (p < 0.001) during various watering regimes. The mean values of water intake were found to be 82.94 ± 1.34 L higher in G3 than G1 and G2.

Use of multivariate adaptive regression splines for prediction of body weight from body measurements in Marecha (Camelus dromedaries) camels in Pakistan.

The aim of the present work was to predict live body weight by means of some body measurements, i.e., SH, CG, and BG in indigenous Marecha camel breed. For this purpose, multivariate adaptive regression splines (MARS) algorithm was used at proportions of various training and test sets, i.e., 65:35, 70:30, and 80:20 in V-tenfold cross-validation. In prediction of live body weight of the Marecha camels (160 female and 145 male animals) in the MARS predictive models, pairs of sex-SH (model 1), sex-CG (model 2), and sex-BG (model 3) as potential predictors. The best MARS model in LW prediction was obtained using sex and SH independent variables for 80:20 training and test set. Sex was determined to be an important source of variation in SH, CG, and BG as a result of sexual dimorphism in camels (P < 0.01). MARS results indicated that SH could be used as an indirect selection criterion to obtain elite camel herds on LW of Marecha camels. If genetically confirmed, the Marecha camels whose SH is taller than 165.1 cm could be selected for providing genetic progress in LW. In conclusion, use of MARS algorithm may be worthy of consideration for better identification of camel breed standards and selection of superior Marecha camels for meat productivity in Pakistan.

Prediction of live body weight based on body measurements in Thalli sheep under tropical conditions of Pakistan using cart and mars.

Thalli sheep is a significant breed reared under tropical region of Punjab province of Pakistan. The present study was conducted to predict live body weight (LBW) by means of from some body measurements, i.e., chest girth (CG), belly girth (BG), rump height (RH), withers height (WH), neck girth (NG), and body length (BL) taken from 155 Thalli indigenous sheep of Pakistan. Age factor is determined to be a significant source of variation for BL, BG, CG, BG, WH, and NG (p < 0.05). LBW is correlated significantly with BL (0.850), CG (0.825), BG (0.849), RH (0.579), WH (0.547), and NG (0.7760), respectively (p < 0.01). For LBW prediction, CART and MARS data mining algorithms were comparatively used based on ten cross-validation method. Among 185 candidate MARS models with 1-5 degrees of interaction and 2-38 terms, the MARS model with 7 terms and no interaction effect in R software was the best model for LBW prediction on the basis of the smallest cross-validated RMSE value. Also, the optimal CART tree structure was obtained with 9 terminal nodes for the smallest cross-validated RMSE value. MARS algorithm outperformed CART in LBW prediction and explained 90.3 (%) of variability in LBW of Thalli sheep. Results of the optimal CART structure reflected that Thalli sheep with BL > 75 cm, RH > 83 cm, and NG > 55 cm has the heaviest LBW of 72 kg. The optimal MARS model displays that the heaviest LBW can be produced by Thalli sheep with BL > 71.12 cm, BG > 106.68 cm, WH > 76.2 cm, NG > 50.8 cm in 5th age group. In conclusion, it coud be recommended that MARS predictive modeling may enable animal breeders to obtain elite Thalli sheep population and to detect body measurement positively influencing LBW as indirect selection criteria for not only describing breed characterization and developing flock management standards, but also ensuring sustainable meat production and rural development in Pakistan.

Use of MARS algorithm for predicting mature weight of different camel (Camelus dromedarius) breeds reared in Pakistan and morphological characterization via cluster analysis.

Mature weight is a significant trait that can be influenced by age, sex, breed, production system, and climate conditions in camels. In camel breeding, it is essential to describe breed standards of the studied camel breeds as part of morphological characterization and to determine morphological traits positively influencing mature weight within the scope of indirect selection criteria. This study was to find the best one among candidate models in prediction of mature weight from several morphological traits measured for eight camel breeds (Bravhi, Kachi, Kharani, Kohi, Lassi, Makrani, Pishin, and Rodbari) raised under Pakistan conditions. The morphological measurements taken from the camels in the study were birth weight (BW), weaning weight (WW), mature weight (MW), age of ridding (ARD), face length (FL), face width (FW), head length (HL), head width (HW), ear length (EL), ear width (EW), neck length (NL), neck width (NW), hump length (HL), hump width (HuW), heart girth (HG), withers height (WH), body length (BL), fore leg length (FLL), and hind leg length (HLL), respectively. In the prediction of mature body weight as a response variable, the optimal MARS predictive model with 15 terms selected by train function of the caret package produced very high predictive performance without encountering overfitting problem. Goodness of fit criteria were estimated to measure predictive quality of the MARS model using ehaGoF package available in R environment. Morphological characterization of the camel breeds was performed with hierarchical cluster analysis (HCA) on the basis of Euclidean distance-Single linkage. At the first step of hierarchical cluster analysis, the similarity level of Bravhi and Kachi camel breeds was the highest with 85.3569 (%). At the second step, Makrani joined to new cluster of Bravhi and Kachi camels found at the first step, and the similarity level of the new cluster comprising Bravhi, Kachi, and Makrani breeds was found as 84.5562 (%). MW was significantly correlated with BW (0.677), WW (0.536), HL (0.524), HuW (0.529), and ARD (0.375) at P < 0.01, and there was the highest correlation of 0.994 between HHL and FLL (P < 0.01). As a result, it could be suggested that results of MARS modeling may help camel breeders to reproduce the elite camel populations and to describe characteristics associated positively with MW within the scope of indirect selection criteria.

Comparative growth response related to hair mineral analysis in dromedary camel calves.

Background: The dromedary camel plays a significant role in supporting the livelihood of pastoral and agropastoral systems, as well as a source of income for the national economy in arid regions. Aim: The current study was executed to check the comparative growth response in relation to hair mineral status in Marecha camel calves reared under an intensive management system and an extensive management system in Thal desert Punjab, Pakistan. Methods: Twelve male and female Camelus dromedarius calves of almost the same weight and age were divided into two groups of 6 each (3 male and 3 female). The calves of the first group were maintained at the Camel Breeding and Research Station, Rakh Mahni, in a semi-open housing system, while the second group was maintained in available housing under field conditions. Calves in the first group were fed concentrate at the rate of 1 kg/head/day along with gram straw (Cicer arientinum) ad libitum, while calves in the second group were allowed to graze/browse for 10 hours daily along with household refusals, including kitchen wastes. Water was provided twice a day. Impressum's digital weighing scale was used for fortnightly weighing. Data collected on different parameters were subjected to statistical analysis with 2 × 2 factorial arrangements of treatments under a completely randomized design. Results: After the 120-day trial period, the mean body weight and average daily gain of male and female calves were significantly increased (p < 0.05) in IMS as 80.8 ± 2.7, 77.8 ± 2.7 kg and 0.67 ± 0.02, 0.65 ± 0.02 kg/days than EMS as 64.5 ± 2.6, 52.9 ± 2.6 kg and 0.54 ± 0.02, 0.44 ± 0.02 kg/days. Intake of crop residues (p < 0.05) was found to be 6.9 ± 0.45 and 6.4 ± 0.45 kg/days for male and female calves in IMS, respectively, and 3.5 ± 0.23 for male and female calves in EMS, respectively. The conversion index g/kg average daily intake was 97.1, 101.5 and 154.3, 125.7 for male and female calves in IMS and EMS, respectively. Regarding hair mineral status, Ca, Mg, Cu, Zn, Fe, and Mn concentrations were found to be significantly different (p < 0.05) among calf groups in IMS and EMS. Conclusion: This study indicates that wool analysis and management of weight gain in camel calves should be further explored to support increased meat supply in arid regions.

Pregnancy status and thyroid function in semi-intensive-kept Marecha she-camels (Camelus dromedarius): managerial implications.

Pakistani Marecha dromedaries, a distinctive breed not covered in the empirical data of the literature reviewed existing for thyroidal physiology in camels, are subjected to investigate thyroid function in pregnant multiparous (n = 22) and non-pregnant nor lactating (n = 22) females. The function of the thyroid gland in pubescent she-camels has evoked discrepancies among researchers exploring this topic in different breeds evolving in divergent environments. Despite season, age, sex, reproductive status, health disorders, and stress are reported as potential modulating factors, these research studies failed to find statistically significant results supporting their hypothesis. For the current research, serum samples were obtained twice a week during one month from semi-intensive-kept Marecha she-camels and analyzed for triiodothyronine (T3) and thyroxine (T4) concentrations. Feed ingredients and additives were quality assessed to examine their nutrient profile, with special attention to protein content and exogenous calcium administration. Statistical differences in serum thyroid hormone levels were found between gravid and non-gravid females, being pregnant she-camels the subgroup with higher levels (P < 0.05). The genetic background of such phenotypic variability in thyroid metabolism in camels has to be further addressed to adapt husbandry practices for breed-specific requirements. Meanwhile, local management strategies are susceptible of accurate routines for sustainable farming.

Impact of early weaning on constituents and nutritional values of camel milk in modern system.

Background: Camel milk is very suitable for human nutritional requirements, and its composition has similarities to mother's milk. Many scientific researches focusing on the myth of nutritional and therapeutic properties of camel milk have been brought to public attention. Aim: This study aims to clarify the impact of early weaning on constituents and nutritional values of the milk of camels reared under a modern farming system during four months postpartum. Methods: Eight lactating camels and eight calves were selected immediately after calving and were assigned to two equal groups under a semi-intensive system in a farm. In the early weaning group (G1), all the calves were allowed to freely suckle their mother's teats from birth to 30 days postpartum. Afterward, the calves were used to stimulate milk letdown for each of the camels' teats for a few seconds before the milking procedure. While in the late weaning group (G2), the calves freely suckled their mother's teats up to 80 days postpartum. Afterward, the calves were restricted from suckling and were used mainly to stimulate milk letdown for the milking procedure. Collection of milk samples started in the 2nd week of postpartum and continued over biweekly intervals up to 16 weeks to determine major fat, protein, lactose, solid non-fat (SNF), and density percentages. Results: The results revealed significant differences in fat, lactose, and density content between groups. We reported significant increases in the percentages of fat, lactose, protein, and SNF levels, besides fluctuations in density with the advancement of the lactation stage. Conclusion: In the early weaning group, the dams produced milk containing high levels of fat and SNF compared to the lactating dams in the late weaning group.

Prevalence and risk factors for IgG antibodies to Neospora spp. in three types of equids from Southern Punjab, Pakistan.

Equine hosts suffer from neurological disease, congenital infection, and reproductive problems associated with Neospora spp. infection. We conducted a cross sectional study using sera from 631 equids (324 horses, 218 donkeys and 89 mules) from the southern region of Punjab province, Pakistan to determine the prevalence of antibodies against Neospora spp. in this diverse group of equines. Fisk factors associated with seropositivity were evaluated statistically based on equine type, breed, age, husbandry, breeding methods, and reproductive history. Prevalence of antibodies to Neospora spp. was detected using a commercially available competitive ELISA kit. We detected IgG antibodies to Neospora spp. in 23.3% of the equids with prevalence by host being 16.0% in horses, 32.6% in donkeys and 26.9% in 89 mules. Statistically significant (P < 0.05) differences in prevalence were observed among these hosts. Prevalence ranged in breeds of horses from 3.7% in the Morna breed, 12.4% in breeding stock, 26.8% in draughting stock, to 31.4% in unknown breeds. No significant (P > 0.05) difference in prevalence was noted among age groups. Prevalence was significantly (P < 0.05) higher in female equines with a history of abortion than those with no history of abortion. This is the first report documenting Neospora spp. in equines from Pakistan and it provides evidence that Neospora spp. is associated with abortions in female equines in Pakistan.