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Abstract Background The ixodid tick genera Rhipicephalus and Haemaphysalis contain several species of medical and/or veterinary importance, but their diversity in some regions of the world remains under-explored. For instance, very few modern studies have been performed on the taxonomy of these genera on the Arabian Peninsula. Methods In this study, we trapped small mammals in the 'Asir Mountains of south-western Saudi Arabia and collected tick specimens for morphological examination and molecular barcoding, targeting three mitochondrial loci: cox1, 16S rRNA and 12S rRNA. Results We obtained a total of 733 ticks (608 Haemaphysalis spp. and 125 Rhipicephalus spp.) from 75 small mammal hosts belonging to six species. All tick specimens were immature except for nine adults recovered from a hedgehog (Paraechinus aethiopicus). Morphologically, the Rhipicephalus ticks resembled R. camicasi, but the Haemaphysalis ticks showed differences in palp morphology compared with species previously described from Saudi Arabia. Phylogenetic analysis and automatic barcode gap discovery identified a novel clade of Rhipicephalus sp. representing most of the nymphs. This was most closely related to R. leporis, R. guilhoni and R. linnaei. The adult ticks and a small proportion of nymphs clustered with R. camicasi sequences from a previous study. Finally, the Haemaphysalis nymphs formed two distinct clades that were clearly separated from all reference sequences but closest to some African species. Conclusions This apparent high level of tick diversity observed in a single study site of only ~ 170 km2, on a relatively small number of hosts, highlights the potential for the discovery of new tick species on the Arabian Peninsula. Graphical Abstract

Abstract Background The Mongolian jird (Meriones unguiculatus) has long been recognized as a permissive host for the filarial parasite Brugia malayi; however, it is nonpermissive to another filarial parasite, canine heartworm (Dirofilaria immitis). By elucidating differences in the early response to infection, we sought to identify mechanisms involved in the species-specific clearance of these parasites. We hypothesized that the early clearance of D. immitis in intraperitoneal infection of the jird is immune mediated and parasite species dependent. Methods Jird peritoneal exudate cells (PECs) were isolated and their attachment to parasite larvae assessed in vitro under various conditions: D. immitis and B. malayi cultured separately, co-culture of both parasites, incubation before addition of cells, culture of heat-killed parasites, and culture with PECs isolated from jirds with mature B. malayi infection. The cells attaching to larvae were identified by immunohistochemistry. Results In vitro cell attachment to live D. immitis was high (mean = 99.6%) while much lower for B. malayi (mean = 5.56%). This species-specific attachment was also observed when both filarial species were co-cultured, with no significant change from controls (U (9, 14) = 58.5, p = 0.999). When we replicated these experiments with PECs derived from jirds subcutaneously infected with B. malayi, the results were similar (99.4% and 4.72% of D. immitis and B. malayi, respectively, exhibited cell attachment). Heat-killing the parasites significantly reduced cell attachment to D. immitis (mean = 71.9%; U (11, 14) = 7.5, p < 0.001) while increasing attachment to B. malayi (mean = 16.7%; U (9, 15) = 20, p = 0.002). Cell attachment to both species was reduced when larvae were allowed a 24-h pre-incubation period prior to the addition of cells. The attaching cells were identified as macrophages by immunohistochemistry. Conclusions These results suggest a strongly species-dependent response from which B. malayi could not confer protection by proxy in co-culture. The changes in cell attachment following heat-killing and pre-incubation suggest a role for excretory/secretory products in host immune evasion and/or antigenicity. The nature of this attachment is the subject of ongoing study and may provide insight into filarial host specificity.

Background: The ixodid tick genera Rhipicephalus and Haemaphysalis contain several species of medical and/or veterinary importance, but their diversity in some regions of the world remains under-explored. For instance, very few modern studies have been performed on the taxonomy of these genera on the Arabian Peninsula. Methods: In this study, we trapped small mammals in the 'Asir Mountains of south-western Saudi Arabia and collected tick specimens for morphological examination and molecular barcoding, targeting three mitochondrial loci: cox1, 16S rRNA and 12S rRNA. Results: We obtained a total of 733 ticks (608 Haemaphysalis spp. and 125 Rhipicephalus spp.) from 75 small mammal hosts belonging to six species. All tick specimens were immature except for nine adults recovered from a hedgehog (Paraechinus aethiopicus). Morphologically, the Rhipicephalus ticks resembled R. camicasi, but the Haemaphysalis ticks showed differences in palp morphology compared with species previously described from Saudi Arabia. Phylogenetic analysis and automatic barcode gap discovery identified a novel clade of Rhipicephalus sp. representing most of the nymphs. This was most closely related to R. leporis, R. guilhoni and R. linnaei. The adult ticks and a small proportion of nymphs clustered with R. camicasi sequences from a previous study. Finally, the Haemaphysalis nymphs formed two distinct clades that were clearly separated from all reference sequences but closest to some African species. Conclusions: This apparent high level of tick diversity observed in a single study site of only ~ 170 km2, on a relatively small number of hosts, highlights the potential for the discovery of new tick species on the Arabian Peninsula. [ABSTRACT FROM AUTHOR]

Aims: ES‐62 is a well‐studied anti‐inflammatory molecule secreted by L4‐adult stage Acanthocheilonema viteae. We maintain the life cycle of A viteae using Meriones libycus as the definitive host. Here, we investigated whether the full life cycle could be maintained, and functional ES‐62 produced, in a related jird species—Meriones shawi. Methods and Results: Adult worms were produced in comparable numbers in the two species, but very few microfilariae (MF) were observed in the M shawi bloodstream. M shawi ES‐62 produced ex vivo was functional and protective in a mouse model of arthritis. Myeloid‐derived cells from naïve and infected jirds of both species were compared with respect to ROS production and osteoclast generation, and some differences between the two species in both the absence and presence of infection were observed. Conclusions: The life cycle of A viteae cannot be successfully completed in M shawi jirds but L3 stage worms develop to adulthood and produce functional ES‐62. Preliminary investigation into jird immune responses suggests that infection can differentially modulate myeloid responses in the two species. However, species‐specific reagents are required to understand the complex interplay between A viteae and its host and to explain the lack of circulating MF in infected M shawi jirds. [ABSTRACT FROM AUTHOR]

Background: The Mongolian jird (Meriones unguiculatus) has long been recognized as a permissive host for the filarial parasite Brugia malayi; however, it is nonpermissive to another filarial parasite, canine heartworm (Dirofilaria immitis). By elucidating differences in the early response to infection, we sought to identify mechanisms involved in the species-specific clearance of these parasites. We hypothesized that the early clearance of D. immitis in intraperitoneal infection of the jird is immune mediated and parasite species dependent. Methods: Jird peritoneal exudate cells (PECs) were isolated and their attachment to parasite larvae assessed in vitro under various conditions: D. immitis and B. malayi cultured separately, co-culture of both parasites, incubation before addition of cells, culture of heat-killed parasites, and culture with PECs isolated from jirds with mature B. malayi infection. The cells attaching to larvae were identified by immunohistochemistry. Results: In vitro cell attachment to live D. immitis was high (mean = 99.6%) while much lower for B. malayi (mean = 5.56%). This species-specific attachment was also observed when both filarial species were co-cultured, with no significant change from controls (U(9, 14) = 58.5, p = 0.999). When we replicated these experiments with PECs derived from jirds subcutaneously infected with B. malayi, the results were similar (99.4% and 4.72% of D. immitis and B. malayi, respectively, exhibited cell attachment). Heat-killing the parasites significantly reduced cell attachment to D. immitis (mean = 71.9%; U(11, 14) = 7.5, p < 0.001) while increasing attachment to B. malayi (mean = 16.7%; U(9, 15) = 20, p = 0.002). Cell attachment to both species was reduced when larvae were allowed a 24-h pre-incubation period prior to the addition of cells. The attaching cells were identified as macrophages by immunohistochemistry. Conclusions: These results suggest a strongly species-dependent response from which B. malayi could not confer protection by proxy in co-culture. The changes in cell attachment following heat-killing and pre-incubation suggest a role for excretory/secretory products in host immune evasion and/or antigenicity. The nature of this attachment is the subject of ongoing study and may provide insight into filarial host specificity. [ABSTRACT FROM AUTHOR]

Jirds (Meriones) are a mostly desert-adapted genus of gerbils, with a wide geographic range, through which it encounters various climatic conditions, which may influence their morphology. In this study, we quantified cranial morphometric variation both within and among seven jird species (M. meridianus, M. hurrianae, M. crassus, M. tristrami, M. persicus, M. libycus and M. vinogradovi), based on a two-dimensional landmark-based geometric morphometric analysis of 972 specimens, covering their entire geographic distribution. The aforementioned analysis was used to compare sexual dimorphism in size and shape within each species, as well as the relationship between size and shape (i.e. allometry) for each species. Despite greatly overlapping in morphospace (when examined visually), statistical analysis indicates significant differences in both size and shape among the seven examined jird species. UPGMA and CVA both show two main species clusters. Deformation grids indicate that these two clusters differ mostly in the relative size of the tympanic bulla, along with differences in the extent of nasal elongation, and the broadness of the zygomatic arch. Allometric changes in shape were analyzed in all species that show an allometric relationship. Sexual dimorphism in shape and size was detected in only three of the seven jird species. A visual inspection of the data indicates high overlap in shape space, and that male skulls are significantly larger than female skulls. When specimens were divided by sex, we found significant allometry in six of the seven species (for both sexes). A factorial multivariate analysis of covariance (MANCOVA) indicated that even when taking size variation into account, the residual shape variation was also significantly different among the sexes of the examined species. The outcome of this study confirms the presence of cranial variation in the examined jirds, and that the patterns of sexual dimorphism and allometry vary considerably among jird species. [ABSTRACT FROM AUTHOR]

Onchocerciasis, a neglected tropical disease prevalent in western and central Africa, is a major health problem and has been targeted for elimination. The causative agent for this disease is the human parasite Onchocerca volvulus . Onchocerca ochengi and Litomosoides sigmodontis , infectious agents of cattle and rodents, respectively, serve as model organisms to study filarial nematode infections. Biomarkers to determine infection without the use of painful skin biopsies and microscopic identification of larval worms are needed and their discovery is facilitated by an improved knowledge of parasite-specific metabolites. In addition to proteins and nucleic acids, lipids may be suitable candidates for filarial biomarkers that are currently underexplored. To fill this gap, we present the phospholipid profile of the filarial nematodes O. ochengi , O. volvulus and L. sigmodontis. Direct infusion quadrupole time-of-flight (Q-TOF) mass spectrometry was employed to analyze the composition of phospholipids and their molecular species in the three nematode species. Analysis of the phospholipid profiles of plasma or serum of uninfected and infected hosts showed that nematode-specific phospholipids were below detection limits. However, several phospholipids, in particular ether lipids of phosphatidylethanolamine (PE), were abundant in O. ochengi worms and in bovine nodule fluid, suggesting that these phospholipids might be released from O. ochengi into the host, and could serve as potential biomarkers. [ABSTRACT FROM AUTHOR]

The Indian Jird, Meriones hurrianae , is unusual among the family Gerbillidae in not being from desert or sub-desert habitats, and in its ability to endure cold weather. Therefore, the distribution range of the Indian jird has no overlap with other species of Meriones . The habitat of the Indian jird is increasingly fragmented, so that this species is increasingly under threat. The ecology of this species, as well as its morphology, is poorly known. The present study, using a two-dimensional landmark-based geometric morphometric approach, aims to investigate intraspecific variation in skull shape and size of populations of this species. For this purpose, more than 50 skull specimens coming from different localities along the distribution range of this species were analyzed. The results allow us to reject the hypothesis that there would be no significant difference in skull shape and size. For instance, some specimens, such as those from Bandar Abbas (Iran), show significant shape differences compared with others. Differences were also significant for some size characteristics, e.g. cranium size and length, as well as the size of the auditory bulla. The observed morphological variation is discussed in the context of taxonomy and conservation of the Indian Jirds. [ABSTRACT FROM AUTHOR]

Haemonchus contortus is an important pathogen of small ruminants and is therefore a crucially important target for anthelmintic chemotherapy. Its large size and fecundity have been exploited for the development of in vitro screens for anthelmintic discovery that employ larval and adult stages in several formats. The ability of the parasite to develop to the young adult stage in Mongolian jirds (Meriones unguiculatus) provides a useful small animal model that can be used to screen compounds prior to their evaluation in infected sheep. This chapter summarizes the use of H. contortus for anthelmintic discovery, offers a perspective on current strategies in this area and suggests research challenges that could lead to improvements in the anthelmintic discovery process. [ABSTRACT FROM AUTHOR]

A series of 4-oxycoumarin derivatives was synthesized, characterized and evaluated in vitro and in vivo for antifilarial activity against the human lymphatic filarial parasite, Brugia malayi . A majority of the compounds studied showed potent in vitro activity with low IC 50 values in the micro molar (μM) range (0.014–1.73 and 0.0056–0.43) against adult worms and microfilariae, respectively. Compounds 8 and 9 were identified to be the most promising antifilarial candidate molecules exhibiting activity in the nanomolar (nM) range. The IC 50 values for compound 8 were 14 nM and 5.6 nM while for compound 9 were 94 nM and 13 nM, respectively, for adult worm and microfilaria. These two compounds also displayed promising adulticidal activity (74.9 ± 4.8% and 69.4 ± 2.8%, respectively) in the primary rodent (jird) screen. This study also serves as a starting point for investigating structure–activity relationship with different amino substituents. [ABSTRACT FROM AUTHOR]

Jirds (genus Meriones) comprise a group of rodents, of which the biodiversity is still poorly known. Reason for this is that several species of similar morphologies are known to occur sympatrically. In the north-west of Iran, four such species occur: Meriones tristrami, Meriones persicus, Meriones vinogradovi and Meriones libycus, prone to several issues of taxonomical ambiguity. A proper characterization of morphological distinctiveness between these species, in relation to the variation within species, could provide the required information for species diagnosis and identification. As some cranial characters of M. tristrami, M. persicus and M. vinogradovi are quite similar, demarcations of species-specific phenotypic variation have proven to be difficult. To tackle this problem, this study involves a geometric morphometric analysis of skull shape and size, incorporating a large representative sample of these four species, originating from most parts of their natural distribution range (especially for M. tristrami). It is first tested whether M. tristrami can be distinguished from the other sympatric species, and if so, to what degree the species shows a geoclimatic pattern in its skull shape and size when comparing different populations. The shape and size analyses show that M. libycus can be distinguished because of its largest skull and the relatively largest tympanic bulla, and that M. tristrami can be distinguished from the other species. At an intraspecific level in M. tristrami, the Iranian groups (Qazvin and west Iran) do not differ in shape among them, but do so in skull size. They could, however, be distinguished in skull shape from the non-Iranian populations included (Turkey and Jordan). To what degree this continuous data can now be translated into discrete and diagnostic features, useful for taxonomic purposes, remains to be studied. [ABSTRACT FROM AUTHOR]

Abstract: Bioassay guided fractionation of ethanolic extract of the leaves of Bauhinia racemosa led to the isolation of galactolipid and catechin class of the compounds (1–7) from the most active n-butanol fraction (F4). Among the active galactolipids, 1 emerged as the lead molecule which was active on both forms of lymphatic filarial parasite, Brugia malayi. It was found to be better than the standard drug ivermectin and diethylcarbamazine (DEC) in terms of dose and efficacy. [Copyright &y& Elsevier]

Abstract: The present study incorporates the findings on in vitro and in vivo antifilarial activity in the marine sponge, Haliclona oculata using an experimental rodent infection of human lymphatic filarial parasite, Brugia malayi. The in vitro antifilarial action was determined on both adult female worms as well as microfilariae using two parameters viz. adverse effect on motility and inhibition in MTT reduction by the treated adult parasite over control worm. The antifilarial activity could be located in the methanol extract and one of its four fractions (chloroform). Bioactivity guided fractionation of chloroform fraction led to localization of in vitro activity in one of its eight chromatographic fractions. Methanol extract, chloroform fraction and one of the chromatographic fractions revealed IC50 values of 5.00, 1.80, and 1.62μg/ml, respectively when adult B. malayi were exposed to these test samples for 72h at 37°C. Under similar exposure conditions, the IC50 values for microfilariae were 1.88, 1.72 and 1.19μg/ml, respectively. The active test samples were found to be safe revealing >10 selectivity indices (SI) on the basis of cytotoxicity to Vero cells (monkey kidney cells) and therefore selected for in vivo evaluation against primary (adult B. malayi intraperitoneal transplanted jird) and secondary (subcutaneous infective larvae induced mastomys) screens. In primary jird model, the three test samples at 100mg/kg for five consecutive days by subcutaneous route demonstrated significant macrofilaricidal efficacy to the tune of 51.3%, 64% and 70.7% by methanol extract, chloroform and chromatographic fraction, respectively. The three samples demonstrated 45–50% macrofilaricidal activity with moderate embryostatic effect in secondary model at 5×500, 5×250 and 5×125mg/kg by oral route. Chromatographic fraction possessing highest antifilarial action was primarily found to be a mixture of four alkaloids Mimosamycin, Xestospongin-C, Xestospongin-D and Araguspongin-C in addition to few minor compounds. [Copyright &y& Elsevier]

Headbobs are up-down movements of the cranium associated with the use of motion parallax for depth perception. Mongolian gerbils (aka jirds; Meriones unguiculatus) often execute a series of headbobs prior to jumping between surfaces. Gerbils were tested in a jumping stand task and headbobs videotaped under three light levels approximating low daylight, dawn/dusk, and moonlight across a range of distances to target. Headbobs per trial increased linearly with increasing distance to the target platform, whereas headbob frequency (rate of headbobbing pre-jump on the start platform) increased with gap distance up to an intermediate level and then decreased. Overall, gerbils made the most headbobs per trial under the darkest conditions, whereas their headbobbing rate was highest for medium illumination, especially for medium-long gap distances. There was a positive correlation between headbob frequency and volume of the superior colliculus (SC), but no relationship between headbobs and relative size of the temporo-posterior (TP) visual cortex. The results suggest that gerbils employ a specific visuomotor strategy for depth perception differentially under different conditions. We suggest that the deployment of headbobs under specific conditions may be part of an SC-driven vigilant state, of which more rapid sampling of the visual environment using headbobs for depth estimation is one component. Moreover, the findings highlight the importance of considering ecological factors in designing studies of visual behavior and its underpinnings in rodents. [Display omitted] • Gerbils deploy headbobs differentially under different lighting conditions. • Gerbils make more headbobs when they have a greater distance to jump and in the dimmest light. • Gerbils' headbob rate is greatest when they have a moderate distance to jump. • Headbob rate is correlated with the size of the superior colliculus. [ABSTRACT FROM AUTHOR]

Abstract: To test the different sensitivity of rodents of the subfamily Murinae and Gerbillinae, Wagner’s gerbils (Gerbillus dasyurus), Tristram’s jirds (Meriones tristrami) and multimammate rats (Mastomys natalensis) were inoculated with Neospora caninum tachyzoites. Clinical signs of neosporosis appeared in all inoculated animals. Histopathological examination confirmed the presence of tachyzoites in brains, lungs, skeletal muscle, myocardium, liver, in serosa of stomach and intestines, and in vesicular accessory genital glands. An examination of brains by PCR revealed presence of N. caninum DNA in all experimentally N. caninum infected rodents. The susceptibility of Wagner’s gerbils and Tristram’s jirds further proved the high sensitivity of gerbiline rodents to the N. caninum infection. The finding of N. caninum tachyzoites in the vesicular accessory genital glands of the infected gerbils suggests the usefulness of the rodent model for demonstration of N. caninum in the male reproductive system. Moreover, the multimammate rat was a susceptible experimental host to be the first immunocompetent rodent of the subfamily Murinae. [Copyright &y& Elsevier]

SUMMARY Filarial infections are characterized by high IgE antibody responses. So far, it is not clear whether IgE antibodies are involved in protection, pathology or both. We established a bioassay to detect reactive IgE antibodies in jirds infected with the filaria Acanthocheilonema viteae. Sera of A. viteae-infected jirds were used to sensitize rat basophil leukaemia (RBL) cells and degranulation was stimulated by addition of antigens of A. viteae. Reactive IgE responses were detected from 2 weeks post infection (p.i.) and throughout the A. viteae infection. Male antigen triggered the strongest mediator release, followed by female worms, infective larvae (L3) and microfilariae. Separation of male and female antigen indicated that several antigens of both genders are potent allergens. In particular, one male specific allergen of about 550 kDa induced strongest degranulation of RBL cells. In addition, mediator release stimulated by antigen fractions of about 15 kDa was due to filarial cystatin. In conclusion, we describe a convenient in vitro assay to examine IgE mediated responses in jirds. A sex specific filarial protein with high allergenic potential is identified and cystatin is established as a potent allergen of A. viteae. [ABSTRACT FROM AUTHOR]

Abstract. The rate of development of immature fleas, Xenopsylla conformis Wagner and Xenopsylla ramesis Rothschild (Siphonaptera: Xenopsyllidae) was studied in the laboratory at 25°C and 28°C with 40, 55, 75 and 92% relative humidity (RH). These fleas are separately associated with the host jird Meriones crassus Sundevall in different microhabitats of the Ramon erosion cirque, Negev Highlands, Israel. This study of basic climatic factors in relation to flea bionomics provides the basis for ecological investigations to interpret reasons for paratopic local distributions of these two species of congeneric fleas on the same host. Both air temperature and RH were positively correlated with duration of egg and larval stages in both species. Change of humidity between egg and larval environments did not affect duration of larval development at any temperature. At each temperature and RH, the eggs and larvae of X. ramesis did not differ between males and females in the duration of their development, whereas female eggs and larvae of X. conformis usually developed significantly faster than those of males. For both species, male pupae developed slower than female pupae at the same air temperature and RH. Air temperature, but not RH, affected the duration of pupal development. At each humidity, duration of the pupal stage was significantly longer at 25°C than at 28°C: 15.3 ± 1.7 vs. 11.7 ± 1.2 days in X. conformis; 14.1 ± 2.0 vs. 11.5 ± 1.7 days in X. ramesis, with a significantly shorter pupal period of the latter species at 25°C. These limited interspecific bionomic contrasts in relation to basic climatic factors appear insufficient to explain the differential habitat distributions of X. conformis and X. ramesis. [ABSTRACT FROM AUTHOR]

Haemonchus contortus is an important pathogen of small ruminants and is therefore a crucially important target for anthelmintic chemotherapy. Its large size and fecundity have been exploited for the development of in vitro screens for anthelmintic discovery that employ larval and adult stages in several formats. The ability of the parasite to develop to the young adult stage in Mongolian jirds (Meriones unguiculatus) provides a useful small animal model that can be used to screen compounds prior to their evaluation in infected sheep. This chapter summarizes the use of H. contortus for anthelmintic discovery, offers a perspective on current strategies in this area and suggests research challenges that could lead to improvements in the anthelmintic discovery process., (Copyright © 2016 Elsevier Ltd. All rights reserved.)