Mashal Muhammad Almutairi

Background: Amikacin is an effective aminoglycoside antibiotic, but its clinical use is limited by nephrotoxicity, hepatotoxicity, and neurotoxicity. N-acetylcysteine (NAC), a glutathione precursor and antioxidant, may help reduce these side effects. This study evaluated whether NAC attenuated selected amikacin-induced biochemical, inflammatory, and histopathological alterations in mice. Methods: Male Swiss albino mice were divided into four groups: control, amikacin, NAC, and amikacin-NAC, and the behavioral performance was assessed using the light-dark box test. Serum samples were collected for measurements of kidney and liver biomarkers, metabolic markers, and electrolytes (calcium, inorganic phosphate, and uric acid). A multiplex cytokine/chemokine panel was used to assess the effects of amikacin with/without NAC on systemic inflammation. Results: Amikacin-treated mice showed a reduction in the time spent in the light area, and this effect was improved by NAC. Amikacin also altered certain biochemistry parameters, increasing levels of aspartate transferase and albumin; these changes were attenuated by NAC. Amikacin caused marked increases in certain inflammatory cytokines and chemokines, including IL-6, IL-12p70, IL-22, IL-18, CXCL10, and CCL11. Treatment with NAC lowered IL-6, IL-12p70, IL-22, IL-18, CXCL10, and CCL11. The sparse partial least squares discriminant and heatmap analysis of the serum inflammatory profile revealed segregation of the amikacin group from the other three groups. Importantly, the histopathological assays on renal and cerebral cortex showed NAC caused improvements, with well-shaped tissues, attenuating the cortical histopathological alterations observed in animals treated with amikacin. Conclusions: These findings suggest that NAC may have the potential to attenuate certain biochemical, inflammatory, and histopathological alterations associated with high-dose amikacin exposure.

Soft ticks are important hematophagous ectoparasites that are found in all zoogeographical regions of the world. In addition to causing direct economic losses, they are also important as disease causing agents. However, in many countries, including Pakistan, their diversity and associated pathogens remained poorly studied. This study aimed to minimize this knowledge gap by conducting morpho-molecular confirmation of soft ticks and molecular assessment of associated Borrelia in this country. A total of 62 ticks were collected from nesting environment of pigeons in Khyber Pakhtunkhwa, Pakistan. Of these, 24.19% (15/62) were identified as Argas hermanni (males = 9, 60%, females = 6, 40%), 62.90% (39/62) as Argas persicus (males = 17, 43.58%, females = 14, 35.89%, nymphs = 8, 20.51%), and 12.90% (8/62) (all nymphs) as an undetermined Argas species. Twenty-nine ticks, representing all three species, were randomly selected for DNA extraction, and subjected to polymerase chain reaction (PCR) to amplify partial fragments of 16S rDNA and CO1 for ticks, and flaB for the associated Borrelia species. Among these, Borrelia DNA was detected in 4/18 (22.2%) A. persicus ticks (2/8 males and 2/7 females). The 16S rDNA and CO1 sequences of A. hermanni showed maximum 98.17% and 99.30% identities, respectively, with conspecifics from Iran, while the CO1 sequence of A. persicus showed 100% identity with conspecifics from Kenya and Kazakhstan. The 16S rDNA and CO1 sequences of Argas sp. showed 96.48% and 97.33% identities, respectively, with A. persicus from Pakistan and Kenya. The flaB sequence associated with A. persicus showed 100% identity with Borrelia anserina from Brazil and Pakistan. In the phylogenetic trees of ticks, A. hermanni (16S rDNA and CO1) and A. persicus (CO1) from the current study clustered with conspecifics, while Argas sp. formed a sister clade with A. persicus group (16S rDNA and CO1). In the flaB phylogenetic tree, B. anserina from the current study clustered with corresponding species in the relapsing fever group. This study provides the first molecular record of A. hermanni, the first report of A. persicus associated with pigeons, with detection of B. anserina, and also documents a novel lineage closely related to A. persicus, all from Pakistan.

Haemaphysalis is the second largest genus of hard ticks (Acari: Ixodidae), both in terms of diversity and global distribution. Despite their wide occurrence, genetic information on this genus remains scarce. This study investigated the genetic diversity, phylogenetic relationships, and associated pathogens of Haemaphysalis ticks infesting small ruminants across northern Pakistan. A total of 613 ticks were collected from 95 infested goats and sheep. Four species were identified based on morphological and molecular analyses, including Haemaphysalis (Ha.) cornupunctata, Ha. danieli, Ha. montgomeryi, and a distinct Haemaphysalis sp. closely related to Ha. sulcata. Species identification was confirmed using mitochondrial 16 S rDNA and cox1 markers. Molecular confirmation of unidentified Haemaphysalis sp. was detected across multiple districts of northern Khyber Pakhtunkhwa, indicating close relation to Ha. sulcata. Pathogen screening revealed Rickettsia hoogstraalii (gltA and ompB) in Haemaphysalis sp. and Ha. danieli, Theileria sinensis-like (18 S rDNA) in Ha. montgomeryi and Ha. danieli, and Anaplasma bovis (16 S rDNA) in Ha. cornupunctata and Ha. montgomeryi. This study provides the first molecular evidence of T. sinensis-like in Ha. montgomeryi and Ha. danieli, R. hoogstraalii in Ha. danieli and Haemaphysalis sp., and A. bovis in Ha. cornupunctata and Ha. montgomeryi from Pakistan. Phylogenetic analyses supported species-level relationships and revealed substantial genetic divergence within Haemaphysalis. These findings highlight the importance of locality-based molecular surveillance for resolving cryptic diversity and improving understanding of tick–pathogen associations in mountainous regions of Pakistan.

Although ticks are recognized as carriers of zoonotic pathogens, the risks of tick-borne infections associated with dogs have received limited attention. The close interaction between humans and dogs increases the zoonotic potential of pathogens, increasing the urgency of addressing this overlooked vector-borne health risk, especially in developing regions. This study sought to identify tick-borne zoonotic pathogens by collecting ticks from dogs and screening for Ehrlichia species, a group known to pose significant yet often neglected health risks to both humans and animals, particularly in the northern area of Khyber Pakhtunkhwa (KP), Pakistan. Ticks were collected from dogs in the Mardan and Dir Lower districts of Pakistan from June 2024 to May 2025. The collected ticks were morphologically identified and processed for molecular identification using 16S ribosomal DNA (16S rDNA) and cytochrome c oxidase ( cox1 ) sequences for ticks, and 16S rRNA and dsb sequences were used for their associated Ehrlichia spp. Among the 223 dogs observed, 147 were infested with ticks, resulting in an overall prevalence of 66%. Rhipicephalus ticks’ infestation were more common in female dogs (91/113, 80.53%) than in male dogs (56/110, 51%). The occurrence of adult females was highest (173/432, 40%), followed by males (158/432, 36.57%) and nymphs (101/432, 23.37%). Two tick species were identified: Rhipicephalus haemaphysaloides and Rhipicephalus sp., which was identified as a member of the Rhipicephalus sanguineus species complex. This was confirmed by 16S rDNA and cox1 sequences, which showed 99–100% maximum identity. In the phylogenetic trees, Rh. haemaphysaloides clustered with the same species reported from Pakistan, while Rhipicephalus sp. grouped with Rhipicephalus sp. morphotype III from Pakistan and India. Regarding their associated bacterial species, such as Ehrlichia spp., they were detected in both Rhipicephalus sp. and Rh. haemaphysaloides based on 16S rDNA and dsb sequences, which revealed maximum identity with Ehrlichia minasensis and Ehrlichia spp., respectively. These species phylogenetically clustered with the corresponding Ehrlichia species. The high infection rate observed in ticks suggests a significant relationship between companion animals and pathogen vectors. Further studies are necessary to investigate the potential health risks of tick-borne Ehrlichia spp. and their zoonotic implications in the region.

Ticks are serious threats to humans, companion and domestic animals, and wildlife throughout the world. Situated in a subtropical region, Pakistan has favorable climatic conditions for various tick species. In Pakistan, ticks cause large but unknown economic losses to both the livestock sector and dairy industry in the form of animal mortality and reduction in dairy products. There is a scarcity of critical knowledge concerning the morphological descriptions, hosts, and vector potential of ticks in Pakistan. To fill this knowledge gap, our study provides current information on the hosts and vector potential of Pakistani ticks, together with a list of tick species occurring in the country. This checklist comprises 49 tick species, 11 belonging to the family Argasidae, and 38 belonging to the family Ixodidae. Of these, 5 argasids and 23 ixodids are redescribed and illustrated here, based on recent collections. Two species—Argas hermanni and Dermacentor montanus—are reported and redescribed for the first time in Pakistan. The spatial distribution, hosts and tick-borne pathogens known for each species are briefly discussed.

Ticks and tick-borne pathogens are expanding their geographic ranges to novel suitable habitats. Together with the World Health Organization (WHO) and the United Nations (UN), Saudi Arabia’s government has joined efforts to prevent the development of tick-associated pathogens. Here, we investigated the prevalence and diversity of antimicrobial-resistant (AMR) bacteria in ticks parasitizing camels in Al-Jouf province. A total of 60 ticks were sampled and identified as Hyalomma excavatum (n = 41) and Hyalomma dromedarii (n = 19), infesting 11 camels. Altogether 70 bacterial isolates were isolated and subjected to Gram staining, followed by identification using the Vitek 2 compact system. Bacterial isolates consisted of 23 different bacterial species. 68.6% (n = 48) of the total isolates were identified as Gram-positive bacteria, comprising 14 different species, while 31.4% (n = 22) of the total isolates were Gram-negative bacteria, comprising 9 different species. Each collected tick was found positive for at least one bacterial species, however, 9 out of the 70 ticks were found to carry 2 or 3 bacterial species. Antimicrobial susceptibility testings showed that the isolated bacteria exhibited resistance to several clinical antimicrobial agents. The antimicrobial susceptibility profile of Gram-positive bacteria showed that 100% (n = 30) were resistant to benzylpenicillin; 93.3% (n = 28) were resistant to and oxacillin; 56.7% (n = 17) were resistant to clindamycin; 53.3% (n = 16) were resistant to vancomycin; 43.3% (n = 13) were resistant to rifampicin; 40% (n = 12) were resistant to erythromycin and trimethoprim/sulfamethoxazole; 30% (n = 9) were resistant to teicoplanin; 3.3% (n = 1) was resistant to tetracycline. All Gram-positive bacteria were 100% susceptible to linezolid, gentamicin, tobramycin, levofloxacin, moxifloxacin, and tigecycline. Susceptibility testing for Gram-negative bacteria revealed, 75% (n = 12) were resistant to cefoxitin, whereas 68.75% (n = 11) were resistant to ampicillin. 62.5% (n = 10) of the Gram-negative bacteria were resistant to ceftazidime. In addition, 50% (n = 8) were resistant to cephalothin, ceftriaxone, and trimethoprim/sulfamethoxazole; 43.75% (n = 7) were resistant to cefepime; 31.25% (n = 5) were resistant to amoxicillin/clavulanic acid; 6.25% (n = 1) was resistant to nitrofurantoin. However, all Gram-negative bacteria were susceptible to other antimicrobials including piperacillin/ tazobactam, imipenem, meropenem, amikacin, gentamicin, ciprofloxacin and tigecycline. The current study sheds light on the AMR burden in ticks infesting camels in Al-Jouf province.