Epidemiological analysis of anaplasmosis in cattle from Khyber Pakhtunkhwa, Pakistan.

Background and Aim: Anaplasmosis, a tick-borne disease affecting livestock caused by the bacteria Anaplasma, poses a global concern. This study aimed to estimate the prevalence, spatiotemporal variation, and associated risk factors of anaplasmosis in cattle from the Bannu and Lakki Marwat districts of Khyber Pakhtunkhwa, Pakistan. Materials and Methods: This study used 197 cattle exhibiting clinical symptoms of anaplasmosis in natural settings. Microscopic examination was used to estimate the prevalence. Potential risk factors, such as sampling regions and months, gender, breed, and age were studied. Results: The study revealed an overall anaplasmosis prevalence of 19.79%. Bannu district exhibited a higher occurrence at 22.10%, compared to Lakki Marwat district at 17.64%. Young cattle (<2 years) demonstrated a notably higher incidence of anaplasmosis (26.78%) compared to adults (>5 years), which had a prevalence of 12.35% (p < 0.05). Female cattle (22.36%) were more susceptible than male cattle (11.11%). Prevalence peaked in June (45.71%) and was lowest in February (3.57%). Crossbred cattle had a higher prevalence (23.52%) than purebred cattle (11.47%). Conclusion: Anaplasmosis can be effectively controlled using a comprehensive approach encompassing selective breeding for resilience, targeted care of young calves and females, effective tick control during warmer months, consistent use of insecticides, and proactive risk factor management. Raising awareness among farmers through diverse channels, including media, is pivotal to bolster tick-borne disease management strategies.

Microplastic Quantification in Aquatic Birds: Biomonitoring the Environmental Health of the Panjkora River Freshwater Ecosystem in Pakistan.

Microplastic pollution has become a global concern, with potential negative impacts on various ecosystems and wildlife species. Among these species, ducks (Anas platyrhynchos) are particularly vulnerable due to their feeding habits and proximity to aquatic environments contaminated with microplastics. The current study was designed to monitor microplastic (MP) pollutants in the freshwater ecosystem of the Panjkora River, Lower Dir, Pakistan. A total of twenty (20) duck samples were brought up for four months and 13 days on the banks of the river, with no food intake outside the river. When they reached an average weight of 2.41 ± 0.53 kg, all samples were sacrificed, dissected, and transported in an ice box to the laboratory for further analysis. After sample preparation, such as digestion with 10% potassium hydroxide (KOH), density separation, filtration, and identification, the MP content was counted. A total of 2033 MP particles were recovered from 20 ducks with a mean value of 44.6 ± 15.8 MPs/crop and 57.05 ± 18.7 MPs/gizzard. MPs detected in surface water were 31.2 ± 15.5 MPs/L. The major shape types of MPs recovered were fragments in crop (67%) and gizzard (58%) samples and fibers in surface water (56%). Other types of particles recovered were fibers, sheets, and foams. The majority of these detected MP particles were in the size range of 300-500 µm (63%) in crops, and 50-150 µm (55%) in gizzards, while in water samples the most detected particles were in the range of 150-300 µm (61%). Chemical characterization by FTIR found six types of polymers. Low-density polyethylene (LDPE) had the greatest polymer detection rate (39.2%), followed by polyvinyl chloride (PVC) (28.3%), high-density polyethylene (HDPE) (22.7%), polystyrene (6.6%), co-polymerized polypropylene (2.5%), and polypropylene homopolymer (0.7%). This study investigated the presence of microplastics in the crops and gizzards of ducks, as well as in river surface water. The results revealed the significant and pervasive occurrence of microplastics in both the avian digestive systems and the surrounding water environment. These findings highlight the potential threat of microplastic pollution to wildlife and ecosystems, emphasizing the need for further research and effective mitigation strategies to address this pressing environmental concern.

First Report on Microplastics Quantification in Poultry Chicken and Potential Human Health Risks in Pakistan.

Microplastics (MPs) are an emerging environmental health concern due to their widespread occurrence in food sources such as fish, meat, chicken, honey, sugar, salt, tea and drinking water, thereby posing possible risks to human health. This study aimed to observe the existence of MPs in the crop and gizzard of the farm chicken, a significant food source in Pakistan. Twenty-four chicken samples were taken from eight poultry farms across Punjab, Pakistan. A total of 1227 MP particles were found from 24 samples (crop and gizzards) originating from the 8 poultry farms. In all, 429 MP particles were found in 24 chicken crops, with a mean of 17.8 ± 12.1 MPs/crop. In contrast, 798 MP particles were found in 24 chicken gizzards, with a mean of 33.25 ± 17.8 MPs/gizzard. Comparatively larger particles, ranging between 300-500 µm, were more abundant (63%) than other considered sizes (300-150 µm [21%] and 150-50 µm [16%]). Additionally, fragments were the dominant type of shape in both sample types (crop [64%] and gizzard [53%]). The predominant colours of particles extracted from gizzards and crops were yellow (32%) and red (32%), respectively. Chemical characterisation of these particles detected four types of polymers: polyvinyl chloride (PVC) at 51.2%, followed by low-density polyethylene (LDPE) at 30.7%, polystyrene (PS) at 13.6% and polypropylene homopolymer (PPH) at 4.5%. In conclusion, we provide evidence for MPs in the gizzards and crops of farmed chickens which may originate from contaminated poultry feed. Only a few studies have been reported globally to assess MPs ingestion in chickens. The current study is the first report from Pakistan. It could be a valuable addition to support MPs literature to establish a relationship between MPs contamination and intake through the food chain.

Toxoplasmosis-Awareness and Knowledge of Pregnant Women in Rural Areas of Malakand Region, Pakistan.

Background: The current study was carried out between October 2017 and October 2018 to explore knowledge, attitudes, practices, and information sources regarding toxoplasmosis among pregnant women in Malakand region, the northwestern part of Pakistan. The current study was carried out between October 2017 and October 2018. Methods: A structured questionnaire was used to interview the women after taking verbal informed consent. Graphpad version 5 was used to indicate the differences. Significant was considered as a P-value of less than 0.05. This study revealed poor knowledge regarding toxoplasmosis. Results: Overall, 31.2% of the respondents showed good knowledge, and 39.2% showed moderate knowledge. On the other hand, 29.5% of the participants showed poor knowledge about toxoplasmosis. The average knowledge score of pregnant women was 79 ± 12.2, which is considered to be within the scale of good knowledge. Number of children within the pregnant multipara women was significantly associated with knowledge about toxoplasmosis. Pregnant women who measured in number of childbirths within a women showed the highest mean score of 42.3 ± 13.3 with 57 (44.8%) displaying a good knowledge level. Pregnant women with more than one child had significantly higher (<0.0001) knowledge scores compared to women with one child or none child. The majority of pregnant women with one child used the social media, followed by mass media as sources of information about toxoplasmosis. Scientific sources of information were used more commonly by pregnant women with none of the child birth. Conclusion: Pregnant women knowledge regarding toxoplasmosis was poor as compared to attitudes and practices. Health workers and newspapers/magazines were the main sources of information.

Antimicrobial Usage and Detection of Multidrug-Resistant Staphylococcus aureus: Methicillin- and Tetracycline-Resistant Strains in Raw Milk of Lactating Dairy Cattle.

Staphylococcus aureus is a prominent cause of food-borne diseases worldwide. Enterotoxigenic strains of this bacteria are frequently found in raw milk, and some of these strains are resistant to antimicrobials, posing a risk to consumers. The main objectives of this study were to determine the antimicrobial resistance pattern of S. aureus in raw milk and to detect the presence of mecA and tetK genes in it. A total of 150 milk samples were obtained aseptically from lactating cattle, including Holstein Friesian, Achai, and Jersey breeds, maintained at different dairy farms. The milk samples were checked for the presence of S. aureus, and it was detected in 55 (37%) of them. The presence of S. aureus was verified by culturing on selective media, gram staining, and performing coagulase and catalase tests. Further confirmation was performed through PCR with a species-specific thermonuclease (nuc) gene. Antimicrobial susceptibility testing of the confirmed S. aureus was then determined by using the Kirby-Bauer disc diffusion technique. Out of the 55 confirmed S. aureus isolates, 11 were determined to be multidrug-resistant (MDR). The highest resistance was found to penicillin (100%) and oxacillin (100%), followed by tetracycline (72.72%), amikacin (27.27%), sulfamethoxazole/trimethoprim (18.18%), tobramycin (18.18%), and gentamycin (9.09%). Amoxicillin and ciprofloxacin were found to be susceptible (100%). Out of 11 MDR S. aureus isolates, the methicillin resistance gene (mecA) was detected in 9 isolates, while the tetracycline resistance gene (tetK) was found in 7 isolates. The presence of these methicillin- and tetracycline-resistant strains in raw milk poses a major risk to public health, as they can cause food poisoning outbreaks that can spread rapidly through populations. Our study concludes that out of nine empirically used antibiotics, amoxicillin, ciprofloxacin, and gentamicin were highly effective against S. aureus compared to penicillin, oxacillin, and tetracycline.

GC-MS Profiling and Biomedical Applications of Essential Oil of Euphorbia larica Boiss.: A New Report.

The present study explored Euphorbia larica essential oil (ELEO) constituents for the first time, obtained via hydro-distillation by means of Gas Chromatography-Mass Spectrometry (GC-MS) profiling. The essential oil was screened in vitro against breast cancer cells, normal cell lines, α-glucosidase, carbonic anhydrase-II (CA-II), free radical scavenging and in vivo analgesic and anti-inflammatory capabilities. The GC-MS screening revealed that the ELEO comprises sixty compounds (95.25%) with the dominant constituents being camphene (16.41%), thunbergol (15.33%), limonene (4.29%), eremophilene (3.77%), and ß-eudesmol (3.51%). A promising antidiabetic capacity was noticed with an IC50 of 9.63 ± 0.22 µg/mL by the ELEO as equated to acarbose with an IC50 = 377.71 ± 1.34 µg/mL, while a 162.82 ± 1.24 µg/mL inhibition was observed against CA-II. Regarding breast cancer, the ELEO offered considerable cytotoxic capabilities against the triple-negative breast cancer (MDA-MB-231) cell lines, having an IC50 = 183.8 ± 1.6 µg/mL. Furthermore, the ELEO was also tested with the human breast epithelial (MCF-10A) cell line, and the findings also presumed that the ELEO did not produce any damage to the tested normal cell lines. The ELEO was effective against the Gram-positive bacteria and offered a 19.8 ± 0.02 mm zone of inhibition (ZOI) against B. atrophaeus. At the same time, the maximum resistance with 18.03 ± 0.01 mm ZOI against the fungal strain Aspergillus parasiticus was observed among the tested fungal strains. An appreciable free radical significance was observed via the DPPH assay with an IC50 = 133.53 ± 0.19 µg/mL as equated to the ABTS assay having an IC50 = 154.93 ± 0.17 µg/mL. The ELEO also offered a substantial analgesic capacity and produced 58.33% inhibition in comparison with aspirin, a 68.47% decrease in writhes, and an anti-inflammatory capability of 65.54% inhibition, as equated to the standard diclofenac sodium having 73.64% inhibition. Hence, it was concluded that the ELEO might be a natural source for the treatment of diabetes mellitus, breast cancer, analgesic, inflammatory, and antimicrobial-related diseases. Moreover, additional phytochemical and pharmacological studies are needed to isolate responsible chemical ingredients to formulate new drugs for the examined activities.

Aroma Profile and Biological Effects of Ochradenus arabicus Essential Oils: A Comparative Study of Stem, Flowers, and Leaves.

The present analysis explores the chemical constituents and determines the in vitro antimicrobial, antidiabetic, and antioxidant significance of the essential oils (EOs) of the stem, leaves, and flowers of Ochradenus arabicus for the first time. The EOs of the flowers presented seventy-four constituents contributing to 81.46% of the total EOs, with the major compounds being 24-norursa-3,12-diene (13.06%), 24-norursa-3,12-dien-11-one (6.61%), and 24-noroleana-3,12-diene (6.25%). The stem EOs with sixty-one compounds contributed 95.95% of the total oil, whose main bioactive compounds were (+)-camphene (21.50%), eremophilene (5.87%), and δ-selinene (5.03%), while a minimum of fifty-one compounds in the leaves' EOs (98.75%) were found, with the main constituents being n-hexadecanoic acid (12.32%), octacosane (8.62%), tetradecanoic acid (8.54%), and prehydro fersenyl acetone (7.27%). The antimicrobial activity of the EOs of O. arabicus stem, leaves, and flowers was assessed against two bacterial strains (Escherichia coli and Streptococcus aureus) and two fungal strains (Penicillium simplicissimum and Rhizoctonia solani) via the disc diffusion assay. However, the EOs extracted from the stem were found effective against one bacterial strain, E. coli, and one fungal strain, R. Solani, among the examined microbes in comparison to the standard and negative control. The tested EOs samples of the O. arabicus stem displayed a maximum potential to cure diabetes with an IC50 = 0.40 ± 0.10 µg/mL, followed by leaves and flowers with an IC50 = 0.71 ± 0.11 µg/mL and IC50 = 10.57 ± 0.18 µg/mL, respectively, as compared to the standard acarbose (IC50 = 377.26 ± 1.20 µg/mL). In addition, the EOs of O. arabicus flowers had the highest antioxidant activity (IC50 = 106.40 ± 0.19 µg/mL) as compared to the standard ascorbic acid (IC50 = 73.20 ± 0.17 µg/mL) using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. In the ABTS assay, the EOs of the same sample (flower) depicted the utmost potential to scavenge the free radicals with an IC50 = 178.0 ± 0.14 µg/mL as compared with the ascorbic acid, having an IC50 of 87.34 ± 0.10 µg/mL the using 2,2-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic acid (ABTS) assay. The EOs of all parts of O. arabicus have useful bioactive components due to which they present antidiabetic and antioxidant significance. Furthermore, additional investigations are considered necessary to expose the responsible components of the examined biological capabilities, which would be effective in the production of innovative drugs.

Risk factors associated with intestinal pathogenic parasites in schoolchildren.

Diseases caused by intestinal parasites impose a substantial burden on population of middle income countries including Pakistan. This research was aimed to assess the risk factors for intestinal parasites in school children of Malakand, Pakistan. Two hundred and eighty eight students were enrolled between February and June 2016. Out of the total enrolled 184 were agreed to collect stool specimens. A questionnaire was also used to collect the data on socio-demographic and socio-economic characteristics of the participants. All the students were guided to collect at least 10gof their own stool specimens. Each of the stool specimens was diagnosed for the presence of any stage of helminth or protozoal parasites. Formal ether concentration method and wet mount techniques were applied. One way ANOVA was used for calculation of P value when it was less than 0.05 which was considered significant. Eighty two percent of the participants were found infected with one species of parasite while 69.9% of the participants were infected with more than one species of intestinal parasites. The most prevalent parasite was hook worm 33.4% (n = 99/296) followed by Taenia saginata 28.7% (n = 85/296), Ascaris lumbricoides 27.7% (n = 82/296), Hymenolepis nana 6.08% (n = 18/296), Entamoeba histolytica 3.37% (n = 10/296) and least for each Enterobius vermicularis and Fasciola hepatica 0.37% (n = 1/296). Previously used drugs, level in school, ages, weight and upper arm circumference were the most significantly (P < 0.05) related factors for the occurrence of intestinal parasite infection. Present research endorsed that risk factors play a key role in the transmission of parasitic diseases. Lack of safe water supply, using raw vegetables, animal keeping, which should be considered for sustainable strategies in the control of these infections preferably in remote parts of the world.

Population diversity of sheep bot fly, Oestrus ovis (Linné, 1758) (Diptera: Oestridae: Oestrinae), using SDS polyacrylamide gel electrophoresis.

The study was planned to evaluate the inter, and intra population genetic variation in general protein banding pattern in Oestrus ovis larvae, by using 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE). The larvae were collected from slaughtered goats head from five different locations (AAS, PN, LA, GM, and BC) of Karachi, Pakistan. The data obtained was subjected to POPGENE (Population Genetic Analysis) software for analysis. The polymorphic loci within populations ranged from 45.45% to 90.91%. Polymorphic loci observed in all populations were 90.91%. The expected heterozygosity observed was 0.182 ± 0.096 in all populations. The chi-square test showed 5 out of 11 loci at H-W equilibrium. The overall fixation index (FST) value was 0.108, showing that the likelihood of subpopulations being differentiated from one another is about 11 percent. The gene flow value (Nm = 2.065) was higher, showing that genes flow occurs between populations. The values of genetic identity were greater, and genetic distance were smaller among all the populations, which means that all the populations were more alike and closer to each other. It was concluded that there was no sympatric and parapatric population differentiation observed among all the population of O. ovis and the populations of the five different locations were not genetically and reproductively isolated from each other.

Changes in the Prevalence of Natural Paramphistomum Cercariae Infection in Indoplanorbis and Lymnaea Intermediate Hosts Influenced by Meteorological Factors.

Paramphistomosis is a neglected ruminant parasitic disease caused by trematodes known as Paramphistomum, which has a diheteroxenic life cycle involving freshwater snail genera, i.e., Planorbis and Lymnaea as the intermediate host and mammals as the definitive host. Snail vector distribution, infection with Paramphistomum spp. cercariae, preferred habitat, and their relationship with certain meteorological factors were not investigated in the province Khyber Pakhtunkhwa of Pakistan. Therefore, this study is designed to evaluate the effects of meteorological factors on the occurrence and severity of Paramphistomum spp. cercariae in Indoplanorbis and Lymnaea intermediate snail hosts. For this purpose, a cross-sectional survey was conducted from October 2018 to September 2019, and snails were collected and then identified using snail shell morphology; their infection with Paramphistomum spp. cercariae was determined through microscopy; and descriptive statistics were used to estimate the prevalence of infection and evaluate their occurrence relationship with a certain meteorological factors including temperature, humidity, rainfall, and pan evaporation in different districts of the above-mentioned province of Pakistan, i.e., adjacent areas of Bara and Kabul rivers in district Nowshehra, Kabul River (Sardaryab) of district Charsadda, Kalpani River of district Mardan, and Indus River (Hund) of district Swabi. A total of 2,706 Indoplanorbis (1539) and Lymnaea (1167) snails were collected, in which overall 10.30% shed Paramphistomum spp. cercariae. The highest infection rate was found in the river of district Swabi (13.20%), while the lowest in adjacent rivers of district Nowshehra (8.19%). Meteorological factors play an essential role in the causation of Paramphistomum spp. infection, parasitic reproduction, vector growth, and survival. Due to these factors, high significant prevalence was found in the summer season (11.83), followed by autumn (11.25), which might be due to optimum temperature, relative humidity, and rainfall (p < 0.05). It was concluded from the study that meteorological factors contribute to the prevalence of Paramphistomum species in the Indoplanorbis and Lymnaea, which act as vectors for the disease paramphistomosis, which may lead to the increased intensity of infection outbreaks of the parasite population in humans and domestic animals.

Prevalence and risk factors assessment of mange mites in livestock of Malakand Division, Pakistan.

Mange mites represent a serious problem in livestock farming due to their worse effect on animals health, reducing milk and meat production. The status of mange mites prevalence was unknown from the livestock present in Malakand Division, Pakistan. Hence, the present study was conducted to determine the prevalence and risk factors assessment of mange mites in livestock of Malakand Division, Pakistan. Villages and livestock herds were visited for data collection and sampling for one year from September 2017 to August 2018. The diagnosis of mange infestation was carried out from skin scrapings collected from symptomatic animals. A total of 1437 animals were screened (240 sheep, 658 cows, 340 buffalos, 199 goats) from selected localities of the Malakand Division. The overall prevalence was 2.37%, with the highest prevalence in buffalos (5.0%) and lowest (0.42%) in sheep (p < 0.05). The locality also significantly affect the prevalence of mange mites in livestock, with a higher percentage (8.14%) in the Ouch area. The prevalence was higher in winter (3.78%) than in summer (1.07%). The prevalence was high (p < 0.05) in young animals (≤1 year) (4.7%) as compared to adults (>1 year) (1.15%). Gender was also a significant risk factor for mange infestation. The infestation was higher in females (2.85%) as compared to males (0.64%) (p < 0.05). Herd size of less than ten animals had a significantly high prevalence rate (3.69%) compared to >10 animals (1.89%). Animals having an infection history >15 days infected more than animals having infection history ≤15 days. This study documented for the first-time epidemiology of mange mites in livestock of Malakand Division, Pakistan. Further studies involving molecular characterization of mange mites will increase our understanding on the spread of mange mites in Malakand Division livestock.

Occurrence of cestodes and comparative efficacy of Typha angustata and sulphadimidine against cestodes in Columba livia domestica (Domestic Pigeon).

The occurrence of intestinal parasites of Columba livia domestica has been on the increase, leading to high economic and production losses with more fatal cases. This study was designed to investigate the prevalence of cestodes in pigeons and determine the efficacy of Typha angustata extract and sulphadimidine against these cestodes in the domestic pigeon. A total of 30 pigeons were examined. 18 (60%) pigeons were found infected with only one type of cestode species (Raillietina spp.). The difference in prevalence between males and females was statistically significant (χ2 = 8.167, p = 0.004). The mean EPG count in group A (treated with T. angustata extract) before treatment and after treatment was 176 ± 4.33 and 155 ± 4.24, respectively. In group B (treated with sulphadimidine), the mean EPG calculated before treatment and after treatment was 184 ± 6.74 and 35 ± 3.53, respectively. The efficacy at day 28 of T. angustata and Sulphadimidine was 11.93% and 80.97%, respectively. It was concluded on the basis of the EPG and efficacy data that T. angustata extract had low efficacy against raillietiniasis, while as sulphadimidine, which is also used before to treat different intestinal parasites, had a good efficacy against raillietiniasis. Further studies are required to know the prevalence of other gastrointestinal parasites in pigeons and efficacy of different medicinal plants against such parasites.

Zoonotic and non-zoonotic helminths in black rats of rain-fed and irrigated areas of Swat, Khyber Pakhtunkhwa, Pakistan.

Present study was conducted to get information on helminth parasites of zoonotic importance among the black rats of district Swat, Pakistan. Two hundred and sixty nine rats were captured from agricultural ecosystem of the district using live captured traps from 2011 to 2013. Captured rats were anesthetized and surveyed for the presence of ectoparasites, then were carefully dissected for investigation of endoparsites. Helminth parasites of 8 species were identified. Presence of parasite was noticed in 23.7% of sampled rats. The infection rates of sampled rats was given in order of their infectivity as Syphacia obvelata 13(4.83%), Aspiculuris tetraptera 13(4.83%), Heterakis spumosa 12 (4.46%), Hymenolepis spp. 9(3.34%), H.diminuta 8(2.97%), Hymenolepis fusa 4(1.48%), Lutziella microacetabularae 4(1.48%) and Lutziella spp. 1 (0.37%). No significant difference (P < 0.4289) was found in prevalence of parasites among areas, crops, crop stages and sex of the host while adult rats were found more infected than sub-adults. S. obvelata and A. tetraptera were the most common species of helminths while Lutziella sp., 1 (0.37%) was found only in one host. Rattus rattus (the black rat) was regarded as the host of helminth parasites of zoonotic importance, therefore the hidden health hazards of this rodent species needed to be considered to prevent infectivity of humans. Current study was concluded that Rattus rattus harbored a wide variety of helminth parasites which shows a hidden risk to inhabitants of the region. Monitoring rats' population in settle areas and educating the local community about the risk of rat borne parasitic diseases transmission through rats appears to be absolutely essential.

COVID-19 pandemic: Mechanistic approaches and gender vulnerabilities.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is a highly pathogenic virus that causes coronavirus-19 disease (COVID-19), a severe respiratory damaging syndrome with serious health complications worldwide. SARS-CoV-2 was unfamilar before the epidemic started in Wuhan, China, in December 2019. COVID-19 is currently a pandemic influencing several countries worldwide. One of the mysteries of the new coronavirus is that it is deadlier for men than women with the male mortality rate is twice as high as that of females.