Cryptosporidium spp are important intestinal protozoan parasites that cause diarrhea in humans, domestic and wild animals. Its infection remains a main public health concern however, the epidemics in human being is still unclear, particularly in developing countries. There are several factors that may enhance the spreading of this parasite in human population especially in young children. A questionnaire was designed to obtain the demographic and clinical data from the participants. A total of 425 stool samples were collected from suspected children (aged 3–10 years) in different hospitals and villages. The initial screening was performed with modified Ziehl Neelsen (mZN) staining technique followed by polymerase chain reaction (PCR). Several potential risk factors were also assessed through the obtained information from suspected individuals. Out of all 425 collected samples, 127 were observed positive by mZN with a prevalence of 29.88% (127/425). The 127 mZN positive samples together with 50 mZN negative samples were processed for molecular analysis through PCR assay. Among them, 71 out of 127 mZN positive samples and 4 out of 50 mZN negative samples were found positive by PCR. The molecular analysis showed that Cryptosporidium parvum was the main cause of infection in children. The results revealed that individuals exposed to diarrhea were more likely to be infected with Cryptosporidium infection while several environmental factors may also play a key role in spreading of this parasite. The current high prevalence of Cryptosporidium infection may be due to the lack of awareness and routine based testing in identification of this parasite in District Buner. Further studies are required to determine the importance of Cryptosporidium infection in this area as well as across the country and to find out the possible risk factors that may be associated with the occurrence of this protozoan. There is, however, an urgent need for laboratory-based observational studies to develop a more dynamic estimate of the cryptosporidial disease burden in the region.

The chitin and chitosan biopolymers are extremely valuable because of their numerous industrial and pharmacological uses. Chitin and chitosan were extracted from the exoskeleton of Periplaneta americana (cockroaches) and termites using various acid and alkali techniques. The extraction process involves an initial demineralization step, during which integument dry powder was subjected to 500 mL (2.07 mol/L) of concentrated HCl at 100 degrees Celsius for 30 min, followed by meticulous rinsing with distilled water to restore the pH to its baseline. Deproteinization was conducted at 80 degrees Celsius using 500 mL (1 mol/L) of NaOH solution, which was repeated for 24 h. A total of 250 mL (0.06 mol/L) of NaOH was added at 100 degrees Celsius for 4 h to obtain chitosan, followed by extensive washing and subsequent drying. FTIR analysis was used to identify the functional groups in Periplaneta americana and termites. The crystallinity of these biopolymers, which have a face-centered cubic structure, was determined by X-ray diffraction analysis. This study assessed the analgesic properties of chitin and chitosan via an acetic-acid-induced writhing test in mice, revealing a significant reduction in writhing behavior following the chitin and chitosan extract. Notably, chitin exhibits the highest degree of analgesic activity compared to chitosan. Both chitin and chitosan show anti-inflammatory effects, with chitosan absorbing proton ions at sites of inflammation, while chitin effectively inhibits ear edema and elicits an analgesic response in mice. Furthermore, the present study revealed antipyretic activity, with termite chitin demonstrating the most significant effect at a concentration of 500 µL/mL, followed by chitosan and chitin at 100 µL/mL. These findings indicate the potential of using chitin and chitosan derived from termites and Periplaneta americana as natural anti-inflammatory compounds, implying prospective uses in anti-inflammatory, antipyretic, and analgesic capabilities.

Rhynchophorus ferrugineus commonly known as Red Palm Weevil is a worldwide distributed and destructive pest of date palm (Phoenix dactylifera) belonging to the family Curculionidae and order Coleoptera. In the present study a total of 365 trees were inspected for Red Palm Weevil infestation in 15 date palm growing areas in district Bannu during September 2013 to June 2014. 500 adults were obtained by capturing alive and dead adult from trees, cocoons and larvae rearing to analyze the size of different body parts and prothoracic black spots and determine the differences and similarities between male and female. Results showed that, 203 trees were infected, and the female was larger in size than the male. Total 13 typologies of prothoracic spots were recorded, among these nine were recorded in both sexes and four only in females. It was concluded that Red Palm Weevil has high diversity worldwide.

Background: Amoebiasis is an intestinal disease caused by enteric protozoan called Entamoeba histolytica belongs to the Genus Entamoeba . The main reason of infection is the contamination of food and water due to the poor sanitation. Among Entamoeba species , Entamoeba histolytica is highly pathogenic while the other species are non-pathogenic and needs no medical treatment. Methodology: A total of 400 stool samples were collected from different areas of district Swat and were processed for screening of amoebic cells. Microscopically identified samples containing amoebic cells were stored at −20 °C till DNA extraction. Extracted DNA was used in a PCR reaction with specific reference primers to amplify the target DNA. Results: Out of all 400 stool samples 111 (27.7%) were found positive through microscopy while PCR reaction confirmed 80 out of microscope positive samples. Among 80 PCR positive samples, the infection with Entamoeba dispar was most common (57.5%) followed by E. histolytica ( 47.5%) and Entamoeba moshkovskii (20%). The positive cases for mono-infection of E. dispar were 33 (41.25%), followed by E.histolytica 25 (31.25%) and E. moshkovskii 7 (8.75%). The co-infection of E. histolytica with E. dispar and E. moshkovskii was 6 (7.5%) and 2 (2.5%), respectively. Similarly the co-infection of Entamoeba dispar with Entamoeba moshkovskii was also 2 (2.5%) while 5 (6.25%) samples were observed with mixed infection of E. histolytica, E. dispar and E. moshkovskii . Significance of the study: The aim of the study was to detect and differentiate the E. histolytica, Entamoeba dispar and Entamoeba moshkovskii using conventional microscopy and polymerase chain reaction. The results suggested that the use of PCR is necessary to differentiate E. histolytica from E. dispar and E. moshkovskii and therefore, to avoid unnecessary treatment the present study recommend the use of PCR for the routine diagnosis of amoebiasis in the study area. It is also suggested that further studies from this area may also facilitate the understanding of genetic diversity of these pathogens.

The GF-1 Soil Sample DNA Extraction Kit is designed for the rapid and efficient purification of bacteria DNA from soil samples without the need for precipitation or organic extraction. The kit uses a high pure specially-treated silica-based material fixed into a column to efficiently bind DNA in the presence of high salt. The kit applies the principle of a minicolumn spin technology and the use of optimized buffers to ensure that only DNA is isolated while cellular protein, humic acid, metabolites, salts and other low molecular weight impurities are removed during the subsequent washing steps. We used this protocol for Cryptosporidium spp DNA extraction from stool samples with minor modifications and its working.

The GF-1 Soil Sample DNA Extraction Kit is designed for the rapid and efficient purification of bacteria DNA from soil samples without the need for precipitation or organic extraction. The kit uses a high pure specially-treated silica-based material fixed into a column to efficiently bind DNA in the presence of high salt. The kit applies the principle of a minicolumn spin technology and the use of optimized buffers to ensure that only DNA is isolated while cellular protein, humic acid, metabolites, salts and other low molecular weight impurities are removed during the subsequent washing steps. We used this protocol for Cryptosporidium spp DNA extraction from stool samples with minor modifications and its working.