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.

Microplastics in water, sediments, and fish at Alpine River, originating from the Hindu Kush Mountain, Pakistan: implications for conservation.

Microplastics (MP) pollution is an emerging threat to life and the environment. These particles are not restricted to human-inhabited lands but also found in different mountains and glaciers where the human population is relatively low. These MPs make their way to the river ecosystem from glaciers, rains, and municipal and industrial effluents. The current study was designed to highlight MPs' pollution in water, sediments, and fishes of the Swat River: originating from the Hindu Kush Mountain Range. These samples were collected from eight different sites across the river. An average concentration of MPs detected in water samples (305.79 ± 289.66 MPs/m3), fish (12.54 ± 8.02 MPs/individual), and sediments (588.29 ± 253.95 MPs/kg). The highest concentration was observed among water samples at Mingora city and the lowest at the confluence point of the rivers near Charsadda being 753.71 ± 330.08 MPs/m3 and 57.64 ± 31.98 MPs/m3, respectively. MP concentrations in the sediment samples were also the highest at Mingora city (834.0 ± 367.21 MPs/kg), and lowest at Chakdara (215.0 ± 20.0 MPs/kg). Among the fish samples, Schizothorax plagiostomus contained the highest while Wallago attu contained the lowest MP concentrations corresponding to 17.08 ± 8.27 MPs/individual and 5.0 ± 2.36 MPs/individual, respectively. Fibers were the most prevalent MPs in all the matrices representing 80%, 92%, and 85% of the total MP count in water, sediments, and fish samples. These findings highlighted that freshwater ecosystem are not free from MPs and are as much vulnerable to anthropogenic activities as marine ecosystem. Therefore, need attention not less than marine ecosystem awareness, education, ecotourism, sustainable reduction in plastic use, and strict rules and regulations could be helpful to prevent the anthropogenic menace.

Effect of dietary supplementation of Neem, Azadirachta indica leaf extracts on enhancing the growth performance, chemical composition and survival of rainbow trout, Oncorhynchus mykiss.

Rainbow trout Oncorhynchus mykiss has a great nutritional value and delicious taste. A 90-days experimental trial was conducted to investigate the effect of dietary leaf extract of neem tree Azadirachta indica as a feeding supplement on the growth performance and proximate composition of O. mykiss. Four experimental diets were designed as T1 (with 5% A. indica leaf extract), T2 (with 7% of A. indica leaf extract), T3 (with 10% A. indica leaf extract), and T4 (control group feed with a regular diet with 0% A. indica leaf extract). The average initial weight of fry 0.4 ± 0.14 g was stocked at 25 fish/tank with two replicates per treatment (4 × 2 = 8). After 90 days of the experimental trial, One-way ANOVA showed significant differences in final body weight, weight gain, specific growth rate, feed conversion ratio, and survival rate among the treatment groups (p < 0.05). The highest final body weight (48.10 g) and weight gain (47.70 g) was observed in T2 with 7% A. indica leaf extract, which was significantly different from the other treatments (p < 0.05). The lowest FCR was recorded in T2 (1.90), which was significantly different compared to other treatment groups (p < 0.05). Inclusion of A. indica leaf extract in formulated feed for rainbow trout had significant effects in the hepatosomatic index, viscerosomatic index and Fulton's condition factor (p < 0.05), but there was no significant difference in the survival rate of rainbow trout fry treated with different experimental diets (p > 0.05). The phenomenal regression indicates that 7.5% A. indica inclusion is optimum for best growth performance for rainbow trout under a controlled environment. Thus, the present study suggests that the dietary leaf extract has performed an excellent nutritional supplement by enhancing growth performance and health conditions of rainbow trout in the hatchery conditions.

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.

Elemental composition of three-spot swimming crab Portunus sanguinolentus (Herbst, 1783) shell from the coasts of Sindh and Balochistan, Pakistan.

The present study aimed to investigate the elemental composition of the hard shell of the three-spot swimming crab, Portunus sanguinolentus, collected from the coasts of Pakistan. Thirty crab shells were collected and divided into three groups considering their size. The element detection was performed by energy-dispersive X-ray spectroscopy with scanning electron microscope (SIM/EDAX). The mean concentration of carbon, oxygen, calcium, copper, magnesium, and phosphorus observed in the shell of P. sanguinolentus was 13.63 ± 6.21%, 46.25 ± 12.62%, 34.39 ± 18.33%, 3.19 ± 1.25%, 1.15 ± 0.99%, and 1.39 ± 0.51%, respectively. The concentration pattern of these observed elements in P. sanguinolentus shells was found in the following order: O > Ca > C > Cu > P > Mg. To compare among the three groups of shells, the highest concentration of calcium (54.60%) was obtained in large-sized shell groups, which was significantly different from the other groups (p < 0.05). A high concentration of copper was found to be accumulated in the large-sized (3.55%) and medium-sized (4.21%) shell groups, which was significantly higher than in small-sized shell groups (p < 0.05). In the case of the large-sized shell group, the mean concentrations of magnesium and phosphorus were significantly lower than the medium-sized shell group (p < 0.05). The results indicate that crab shells could act as a good bio-sorbent for several minerals in its ecosystem. P. sanguinolentus shell is very rich in calcium, magnesium, and phosphorus. Extraction of calcium, magnesium, and phosphorus from P. sanguinolentus shells could be profitable for biofertilizer and pharmaceutical industries.

Effect of feeding frequency as a rearing system on biological performance, survival, body chemical composition and economic efficiency of Asian seabass Lates calcarifer (Bloch, 1790) reared under controlled environmental conditions.

A 70-day rearing trial was done to determine the optimal frequency of feeding on growth performance (GP), feed conversion rate (FCR), cannibalism, survival rate (SR), body chemical composition and economic efficiency of the Asian sea bass. This study tested four different treatments of feeding frequencies (FF), once (T1), twice (T2), three times (T3), and four times (T4) per day. An average initial weight of Asian sea bass fry was 0.2 g (SD = ±0.12) were stocked 10 individuals per m3 (9.14 m × 1.82 m × 1.22 m, L × W × H; water depth 0.61 m) with two replicates per treatment (4 × 2 = 8). Fry were fed a mixture of larval commercial feed and shrimp with a pellet diet containing (46% CP). Initially, the feeding rate of 8% biomass per day was further adjusted according to fish biomass on a weekly basis. Results showed that, the FF significantly affected (p < 0.05) on growth indictors and survival rate (SR). Specifically fry fed three times a day (T3) had the best FBW, FL, SGR, ADWG and FCR followed by T4 and T2 while fry fed one time a day was the lowest in these parameters. Also, VSI, HSI and CF (k) significantly differed among the treatments. The fish whole body content of protein, moisture and ash did not significantly (p < 0.05) be affected by feeding frequency, but lipid content differed and both T3, T4 were the highest. It could be concluded that, increasing FF up to three times a day had a positive effect on weight gain, survival rate and feed utilization of Lates calcarifer. The second degree polynomial regression indicates that fed three times a day is optimum for best growth performance and survival for Asian sea bass.

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.