Assessment of Babesia spp. prevalence in various domestic animals across Southern Punjab, Pakistan.

Parasitic diseases, notably babesiosis, exert a substantial impact on the global cattle industry, posing challenges to commerce, economies, and human health. This study, conducted in Southern Punjab, Pakistan, aimed to assess the prevalence of Babesia spp. across various livestock species using microscopic and PCR methods. A total of 180 blood samples (60 from each district) were systematically collected from apparently healthy animals, with 36 samples obtained from each domestic animal species, including camel, cattle, buffalo, goat, and sheep, noting that 12 samples were collected from each district for each animal species. Overall prevalence was determined to be 32.8% (59/180), with varying rates among species: 25.0% in cattle, 41.66% in buffalo, 30.55% in goats, 33.3% in sheep, and 33.3% in camels. Microscopic examination revealed slightly varied infection rates among large and small domestic animals (22.2%), while PCR results indicated a 32.8% overall infection rate in both large and small domestic animals, with no statistical significance. District-wise analysis showed regional variations, with Muzaffargarh recording a prevalence rate of 23.33% through microscopic examination, while Lodhran and Bahawalpur recorded 21.67%. PCR results revealed higher rates (38.33%, 26.67%, and 33.33%, respectively), underlining the importance of employing PCR for accurate detection. Examining ruminant types, large ruminants exhibited a 32.4% infection rate, while small domestic animals showed 33.3%, with no significant difference (p=0.897). District-wise prevalence showcased significant variation, with Muzaffargarh demonstrating a 25% prevalence, Lodhran 22%, and Bahawalpur 22%, through microscopic examination. PCR results displayed 38.33%, 27%, and 33.3%, respectively, with no statistical significance. Detailed analysis of individual districts highlighted variations in infection rates among camels, cattle, buffalo, goats, and sheep. The binomial test indicated significant differences through microscopic analysis (P=0.011) but non-significant variations through PCR (P=0.065), emphasizing the precision of PCR. Regional variations in prevalence, notably with Punjab exhibiting the highest frequency (33.87%) and KPK the lowest (13.24%), suggest potential influences from varying veterinary practices and environmental factors. This study underscores the pivotal role of PCR alongside microscopy for accurate babesiosis diagnosis. These findings contribute to the broader understanding of babesiosis prevalence, emphasizing the necessity of advanced molecular techniques for informed control measures.

Molecular epidemiology, associated risk factors, and phylogeny of Theileria annulata infecting buffaloes and cattle from different agro-climatic regions of Punjab, Pakistan.

Background: Tropical theileriosis is the most prevalent hemoprotozoan disease in Pakistan. Aims: The study aimed to investigate the epidemiology and evolutionary relationship of Theileria annulata in bovines in diverse agro-climatic regions of Punjab, Pakistan. Methods: 800 blood specimens were collected from asymptomatic cattle (n=480) and buffaloes (n=320) using a multistage sampling method from Sargodha (n=400) and Multan (n=400) districts. The samples were assessed for blood smear microscopy and cytochrome b gene based PCR. Twenty samples were collected from each union council of each district. Results: The overall prevalence of T. annulata infection in bovines was 9% and 17.13% as determined by blood smear analysis and PCR, respectively. The disease positivity in cattle and buffaloes was respectively 10.21% and 20.42% by blood smear screening and 7.19%, 12.19% by PCR. The overall PCR based prevalence in the Sargodha and Multan districts was 19% and 15.25%, respectively. Absence of rural poultry, tick infestation, and a history of tick-borne diseases had significant effect in cattle. Tick infestation and age were the main statistically significant disease determinants in buffaloes. The evolutionary analysis of the cytochrome b gene showed that the Pakistani isolate infecting buffalo was related to those from Iran, India, Egypt, and Sudan. The isolate from cattle was genetically close to those from Pakistan, India, Iran, Iraq, and Turkey. Conclusion: It can be concluded that biotic and abiotic factors contribute to disease occurrence. The current study will help to devise control strategies to prevent substantial economic losses.

Phylogenetic analysis of halophyte-associated rhizobacteria and effect of halotolerant and halophilic phosphate-solubilizing biofertilizers on maize growth under salinity stress conditions.

AIMS: The main objective of the present work was to evaluate plant growth-promoting abilities of bacterial strains from the rhizosphere of halophytes and their effect on maize growth under salinity stress. METHODS AND RESULTS: Halophilic bacteria were identified using 16S rRNA sequence analysis and their plant growth-promoting abilities were characterized. Phylogenetic analysis showed that bacterial strains belonging to Bacillus, Halobacillus and Pseudomonas were dominant in the rhizosphere of halophytes. More than 93% strains showed P-solubilization activity and IAA production. About 54% strains were able to produce ACC deaminase, 29% strains showed positive results for nitrogen fixation, 41 and 21% strains showed siderophores and HCN production ability respectively. More than 90% strains showed antifungal activity against more than two fungal pathogens and production of different hydrolytic enzymes. To study the plant growth-promoting effect on maize, five bacterial strains Bacillus safensis HL1HP11 and Bacillus pumilus HL3RS14, Kocuria rosea HL1RP8, Enterobacter aerogenes AT1HP4 and Aeromonas veronii AT1RP10 were used as inoculants; in the form of seed coat and enriched soil-based phosphate biofertilizers. All bacterial strains positively affected the maize growth as compared to non-inoculated control + NaCl plants. Plants inoculated with Bacillus HL3RS14-based soil biofertilizers showed maximum increase in dry weights of root (48-124%) and shoot (52-131%) as compared to control + NaCl (soil + rock phosphate, no inoculum). PGPR inoculations under salinity stress conditions showed high concentrations of proline, glycine betaine and malondialdehyde. CONCLUSION: These results indicated that under saline soil conditions, halophilic PGPR strains combined with carrier materials are promising candidates as biofertilizers.

Genome Sequence of the Mycorrhizal Helper Bacterium Pseudomonas fluorescens BBc6R8.

We report the draft genome sequence of the mycorrhizal helper bacterium Pseudomonas fluorescens strain BBc6R8. This is the first genome of a mycorrhizal helper bacterium. The draft genome contains 6,952,353 bp and is predicted to encode 6,317 open reading frames. Comparative genomic analyses will help to identify helper traits.

Isolation and 16S rRNA sequence analysis of the beneficial bacteria from the rhizosphere of rice.

The present study deals with the isolation of plant growth promoting rhizobacteria (PGPR) from rice (variety NIAB IRRI-9) and the beneficial effects of these inoculants on two Basmati rice varieties. Nitrogen-fixing activity (acetylene-reduction activity) was detected in the roots and submerged shoots of field-grown rice variety NIAB IRRI-9. Estimation of the population size of diazotrophic bacteria by ARA-based MPN (acetylene reduction assay-based most probable number) in roots and shoots indicated about 10(5)-10(6) counts/g dry weight at panicle initiation and grain filling stages. Four bacterial isolates from rice roots and shoots were obtained in pure culture which produced phytohormone indoleacetic acid (IAA) in the growth medium. Among these, three isolates S1, S4, and R3 reduced acetylene to ethylene in nitrogen-free semi-solid medium. Morphological and physiological characteristics of the isolates indicated that three nitrogen-fixing isolates S1, S4, and R3 belonged to the genus Enterobacter, while the non-fixing isolate R8 belonged to the genus Aeromonas. 16S rRNA sequence of one isolate from root (R8) and one isolate from shoot (S1) was obtained which confirmed identification of the isolates as Aeromonas veronii and Enterobacter cloacae, respectively. The 1517-nucleotide-long sequence of the isolate R8 showed 99% similarity with Aeromonas veronii (accession No. AF099023) while partial 16S rRNA sequence (two stretches of total 1271 nucleotide length) of S1 showed 97% similarity with the sequence of Enterobacter cloacae (accession No. AJ251469). The seedlings of two rice varieties Basmati 385 and Super Basmati were inoculated with the four bacterial isolates from rice and one Azospirillum brasilense strain Wb3, which was isolated from wheat. In the rice variety Basmati 385, maximum increase in root area and plant biomass was obtained in plants inoculated with Enterobacter S1 and Azospirillum Wb3, whereas in the rice variety Super Basmati, inoculation with Enterobacter R3 resulted in maximum increase of root area and plant biomass. Nitrogen fixation was quantified by using 15N isotopic dilution method. Maximum fixation was observed in Basmati 385 with the inoculants Azospirillum Wb3 and Enterobacter S1 where nearly 46% and 41% of the nitrogen was derived from atmosphere (%Ndfa), respectively. In general, higher nitrogen fixation was observed in variety Basmati 385 than in Super Basmati, and different bacterial strains were found more effective as inoculants for the rice varieties Basmati 385 and Super Basmati.

A study on the impact of maternal knowledge and practice on the nutritional status of infants.

PIP: In India, health workers interviewed 123 mothers of infants attending the child health clinic of the S.K. Institute of Medical Sciences in Srinagar to determine whether maternal knowledge and practice were associated with the nutritional status of the infants. 28 children were considered to be well nourished, while the remaining 95 children were determined to be in various degrees of malnutrition. Mothers whose infants were well nourished had a higher level of breast feeding knowledge than did those whose infants were moderate to severely malnourished (score, 27.13 vs. 16.01-18.75; p .0001). None of the mothers of malnourished infants had an excellent score on breast feeding practices. Differences in the mean score values for breast feeding practices between all consecutive grades of nutrition were significant (20.22 for excellent score, 16.85, 15.53, and 14.18 for grades I, II, and III, respectively; p .001). The only mothers who had an excellent score for infant weaning awareness were 3 mothers whose infants had an excellent nutritional status. A significant difference in mean score values for knowledge of infant weaning between mothers of well nourished infants and grade I malnourished infants (26 vs. 17.19; p .001) as well as between those of grade II malnourished infants and grade III malnourished infants (17.06 vs. 13.64; p .01) were significant. Little difference in infant nutritional status existed between mothers who scored fair and those who scored poor, but, among mothers of well nourished infants, those who scored well were more likely to be have infants of good nutritional status than those who did not score well (47.05% v. 19.56% and 18.33%; p .001). These findings show a decreasing trend between awareness and practice of breast feeding/infant weaning, suggesting that further improvement of health education is needed to reduce the lag between breast feeding awareness and practice.^ieng