The endogenous life cycle of Eimeria caliginosa was studied in experimentally infected dusky rice rats, Melanomys caliginosus. All endogenous stages were located in the epithelial cells of villi in the small intestine. Both Giemsa-stained mucosal scrapings and histological sections were studied for the diagnosis of all the life-cycle stages. Eimeria caliginosa has 3 generations of meronts (M) that differ by size, shape, and number of merozoites (m), which also differ in their size, shape, and location of their nuclei within the cytoplasm of the meronts. The 3 meront types, M₁–M₃, respectively, had 20–33 (m₁), 5–9 (m₂), and 13–16 (m₃) merozoites. Macrogametocytes and microgametocytes, as well as macrogametes and microgametes, completed the sexual cycle, which concludes with the formation of unsporulated oocysts. This parasite's endogenous development produced severe intestinal lesions in the experimentally infected dusky rice rats. In our ongoing work to understand the biodiversity present in plants and animals of the protected Reserva Biológica Alberto Manuel Brenes (ReBAMB) field station in Costa Rica, we now have discovered 3 new Eimeria species, and this is the second complete life cycle in which we document both the asexual and sexual stages.

Species composition and diversity of dactylogyrids were compared on gills of wild and cultured goldfish (silver crucian carp) Carassius auratus from 3 naturally populated lakes and 3 stocked aquaculture ponds in the Hubei province of China to examine the differences in the gill parasite community between these natural and farmed waters. Of the 7 Dactylogyrus spp. detected, all were found in lakes and 5 in ponds, with Dactylogyrus inexpectatus and Dactylogyrus anchoratus being absent from ponds. No significant correlation was found between the species richness and habitat area or host size, nor was there a significant difference in mean species richness between lakes (0.41–0.65) and ponds (0.30–0.76). Brillouin's diversity in lakes (0.049–0.067) was higher than that in ponds (0.024–0.046), but not significantly so. Although the diversity of parasite communities was higher in wild goldfish, higher mean abundance of some Dactylogyrus spp. was found in cultured goldfish. Based on Bray–Curtis similarity, it was difficult to differentiate parasite communities in lakes from those in ponds at the infracommunity level, whereas the 3 lakes and Guanqiao pond differed markedly from the remaining 2 ponds at the component community level. Although infracommunities differed among waterbodies, no effects of fish length or waterbody type were found on infracommunity or component community structure. Together, these results suggest that abundance and species richness of Dactylogyrus spp. on goldfish in lakes and farm ponds are influenced by habitat-specific environmental factors.

Water mites of the genus Unionicola are the most common symbionts of freshwater bivalves. During the current investigation, a total of 120 live freshwater mussels representing 5 species, Corbicula fluminea (Veneroida), Coelatura aegyptiaca (Unionoidea), Mutela rostrata, and Chambardia rubens (Mutelidae), were collected from 2 localities in Tura (Helwan Governorate) and El Kanater (Qaluobiya Governorate), Egypt. Only 3 of the 4 bivalve species listed are considered freshwater bivalves (members of Unionoidea). Corbicula fluminea belong to the family Cyrenidae within Veneroida. Collected mussels were dissected and examined for the presence of unionicolid mites. It was found that 30.83% (37/120) were infected with a single mite species, Unionicola tetrafurcatus (Unionicolidae). The highest prevalence was observed during the summer with 83.33% (25/30) whereas the least was observed in autumn, i.e., 33.33% (10/30). Mites were recovered from the gills, gonads, and visceral mass of mussel hosts. Gills of host mussels were the primary site of oviposition for Unionicola mites. Smaller bivalves in size had significantly greater numbers of mites than did larger ones in size. Numbers of mites per host species was variable and the highest prevalence level of 83.33% (25/30) was recorded in Cor. fluminea while the lowest one of 16.66% (5/30) was found in Ch. rubens. Morphological and morphometric characterizations of mites revealed some differences between the present species and other related Unionicola. Histopathological responses of host mussels to the eggs, larvae, and cuticular remnants of U. tetrafurcatus were also studied. Therefore, the present study demonstrated that freshwater bivalves have a new host and locality records for infection with U. tetrafurcatus. Future studies are recommended to include advanced molecular characteristics for these mites.

Laelapinae mites are common ectoparasites of small mammals, mainly sigmodontine rodents, but males are not frequent in natural populations. In this study, the morphology of the unknown males of Androlaelaps misionalis Lareschi, 2010 and Androlaelaps ulysespardinasi Lareschi, 2011 are described. The males of these species differ from the male of Androlaelaps rotundus (da Fonseca, 1936), a close species, because of the size and shape of their dorsal shield and the length of paranal and sternal st1 and st2 setae. The males of A. misionalis and A. ulysespardinasi differ in their size, in the distance of j6 setae in the dorsal shield, length of hypostomal seta hyp3, number of accessory setae in holoventral shield, length of postanal and sternal st3 setae, number of opisthogastric setae in unarmed integument lateral to level of coxae IV, and shape of tip of movable digit in the chelicera.

The genotyping of 25 isolates of Toxoplasma gondii from free-range chickens in the state of Bahia, Brazil, was performed by PCR-restriction fragment length polymorphism using 11 genetic markers: SAG1, 5′ 3′SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3. The analysis revealed 8 genotypes, 3 of which had not been previously reported. Four genotypes were represented by single isolates, whereas the other genotypes were represented by 2 or more isolates. Five isolates showed mixed infections, and 2 of them were identical. None of the clonal types I, II, or III were found, but 2 isolates corresponded to the Brazilian clonal lineage BrIII. There was a single allele for the c22-8 marker. The CS3 marker demonstrated efficiency in the evaluation of virulence in mice. This study reaffirms the diverse genetic variability of T. gondii in Brazil.

Rodents play an important role in the transmission of Toxoplasma gondii; however, little is known about the seroprevalence and genetic diversity of T. gondii infection in rodents in Yunnan Province, China. In this study, we examined the seroprevalence and genetically characterized T. gondii infection in naturally infected rodents in Yunnan Province between March and July 2016. In total, 261 serum samples were collected from rodents in the Jingha district, Xishuangbanna Dai Autonomous Prefecture, Yunnan Province, and examined for T. gondii antibodies by modified agglutination test (MAT) at a 1:20 cut-off, with titers of 1:20 in 24 samples, 1:40 in 10, 1:80 in 10, and 1:160 in 4. Thirty-two of 261 (12.26%) serum samples were positive for T. gondii by MAT. In total, 751 rodent brain samples were examined by semi-nested PCR; 46 (6.13%) were positive for the T. gondii B1 gene, 2 of which showed complete genotyping results for all 11 polymorphic loci (SAG1, SAG2, alt.SAG2, SAG3, BTUB, GRA6, L358, PK1, C22-8, C29-2, and Apico). The 2 samples were characterized to represent ToxoDB#137 ( http://toxodb.org/toxo/). These findings enriched the epidemiological and genetic diversity of T. gondii in China and have implications for the better prevention and control of T. gondii infection in humans and animals.

The human liver fluke Opisthorchis viverrini (Platyhelminthes, Trematoda, Digenea) uses snails of the genus Bithynia as first intermediate host. Peculiarly among trematodes, the eggs of O. viverrini hatch within the digestive tract of its snail host. It remains uncertain whether hatching in this species is mediated through mechanical fracture of the eggshell or by digestion with specific digestive enzymes. This study aimed to characterize enzymes with specific inhibitors and factors involved in the hatching activity of O. viverrini eggs. For measuring egg hatching in vivo, 50 O. viverrini mature eggs were fed to individual Bithynia siamensis goniomphalos snails at various temperature conditions for 24 hr. Ex vivo, mature eggs were incubated with crude snail extract and commercial leucine aminopeptidase (LAP). Egg-hatching of O. viverrini was temperature dependent, with optimal hatching occurring at 24–28 C, with a peak of hatching of 93.54% in vivo and 30.55% ex vivo occurring at these temperatures. Ex vivo hatching rates increased to 45.87% under anaerobic conditions at 28 C. Some 22.70% and 16.21% of heat-killed eggs also hatched within the snail digestive tract and snail extract, respectively, indicating that host molecules are involved in the hatching response. Most eggs hatch in the anterior regions of the digestive tract. Hatching was completely inhibited in the presence of bestatin, an inhibitor of LAP, but not in the presence of phosphatase inhibitors. Bestatin inhibition of hatching was reversible. Finally, egg hatching could be induced by addition of a porcine LAP. The results indicate that this digenean utilizes both LAP of the snail host and movement of miracidia for hatching.

Samples obtained from 11 teleost fish species collected in waters off of the Mediterranean coasts of Sicily, Italy, were examined for the presence of Hysterothylacium spp. larvae. In total, 3,017 fish samples were examined, and the larvae recovered were used in subsequent phylogenetic studies. Fifty-eight raphidascaridid parasitic nematodes were found in the examined fish, with prevalence values ranging from 0.2% in anchovies to 60% in forkbeard samples. Twenty-seven parasites were identified as Hysterothylacium fabri and Hysterothylacium aduncum by sequencing the following regions: the nuclear ribosomal internal transcribed spacer (ITS) and the mitochondrial cytochrome c oxidase subunit II (COXII). Two other larvae collected from Lophius piscatorius had very low identity scores, with ITS and COXII sequences reported in GenBank and high genetic distances (AY603539). Pairwise comparisons between the ITS region of the H. fabri isolated from fishes from Sicilian coastal waters and those isolated from the Mediterranean Sea, Turkey (KC852206), revealed genetic differences ranging from 0.015 to 0.018. Our H. aduncum samples had very low genetic differences to H. aduncum from the Adriatic Sea (KP979763, 0.00–0.003). The concatenated phylogenetic examination of the ITS-COXII sequences by using maximum likelihood analyses indicated 3 distinct clades supported by high bootstrap values. Further molecular identification and detailed morphological analyses are needed to clarify these results and confirm the diversity and relationships within Hysterothylacium.

Several genes have recently been shown to affect the innate resistance of laboratory strains of Biomphalaria glabrata to infection with Schistosoma mansoni, including Hsp90, the expression of which following a brief exposure to elevated temperature can interfere with resistance in juvenile BS-90 snails. Because a prior study failed to see a similar effect in adult snails, juvenile BS-90 snails were exposed to 10 or 50 miracidia following a 4.5 to 6 hr incubation at 33 or 37 C. Snails were then monitored for production of secondary sporocysts or release of cercariae. In addition, snails exposed to 33 C were examined histologically between 1 and 15 days post exposure (DPE) to 30 miracidia to assess the fate of primary sporocysts. Other than elevated numbers of viable primary sporocysts in the tissues of heat shocked snails at 3 DPE, no statistically significant effect of elevated temperature was observed. This discrepancy with regard to prior studies is hypothesized to result from genetic divergence in different laboratory colonies of the same strain of snail.

Canine hepatozoonosis, caused by Hepatozoon canis, is a tick-borne disease in domestic and wild dogs that is transmitted by ingestion of Rhipicephalus sanguineus ticks. The aim of the study was to detect H. canis in stray dogs in Iran with the use of blood smear examination and molecular techniques. From October 2014 to September 2015, 150 EDTA blood samples were collected from stray dogs in northeastern Iran. Blood smears were microscopically examined for the presence of Hepatozoon gamonts; whole blood was evaluated by PCR with subsequent sequencing and phylogenetic analysis. Hepatozoon spp. gamonts were observed in the neutrophils of 5/150 (3.3%) blood smears, whereas Hepatozoon spp. 18S rDNA was detected in 12/150 (8.0%) blood samples from stray dogs. There was a good correlation between microscopy and PCR methods (κ = 0.756). The highest seasonal rate of infection was detected in the summer (P < 0.05). The frequency of Hepatozoon spp. infection did not differ significantly by gender or age (P > 0.05). The alignment analysis of the sequenced samples showed ≥99% similarity with other nucleotide sequences of Hepatozoon spp. in GenBank. The phylogenetic tree also revealed that the nucleotide sequences in this study were clustered in the H. canis clade and different from the Hepatozoon felis and Hepatozoon americanum clades. Accordingly, we conclude that H. canis infection is present among dogs in northeastern Iran.

Parasitic nematodes affecting the respiratory system of cats are gaining much attention in the scientific community and new knowledge has recently been acquired on their epizootiology, biology, and clinical aspects. In particular, in the past few years different studies have indicated that the metastrongyloid lungworm Troglostrongylus brevior can be vertically transmitted in cats, although information on its basic biology is still poor. The present paper describes a case of troglostrongylosis in a litter of 3 suckling kittens, with a focus on anatomo-pathological and biological features of the infection in 2 of these 3 animals. The third kitten survived the infection and clinical and therapeutic aspects are presented. New insights into the transmission and biology of T. brevior are discussed along with potential strategies for an efficacious treatment and control of this life-threatening parasitosis of cats.

Occurrence of clinical signs by infection with Baylisascaris potosis, the roundworm of kinkajous (Potos flavus), in mice, rats, and rabbit were studied, and the migration behavior of larvae in mice were compared with that of Baylisascaris transfuga, the roundworm of bears (Ursus spp.). Three groups of 8 mice, 3 groups of 6 rats, and 3 groups of 2 rabbits were inoculated with either 10, 100, or 1,000 B. potosis eggs. The other 8 mice were inoculated with 1,000 B. transfuga eggs. Animals were monitored for the occurrence of clinical signs until 60 days postinoculation (DPI). The carcass, viscera, brain, and eyes of each of 6 mice inoculated with 1,000 eggs of B. potosis or B. transfuga at 60 DPI were removed individually, and the number of larvae was counted. One mouse inoculated with 100 B. potosis eggs showed rolling at 27 DPI, and 1 larva was found in the medulla oblongata of this mouse. No clinical signs were observed in the other mice or in the rats and rabbits. A mean of 387.2 larvae was recovered from mice inoculated with 1,000 B. potosis eggs, and a mean of 422.0 larvae from mice inoculated with 1,000 B. transfuga eggs. The highest number of larvae was recovered from the carcasses for both B. potosis and B. transfuga. In the viscera, higher numbers of B. transfuga larvae (mean 131.8) were seen than B. potosis larvae (mean 33.1). In the brain, only 1 larva was detected in 1/6 mice inoculated with 1,000 B. potosis eggs, whereas a mean of 21 larvae was detected in mice inoculated with 1,000 B. transfuga eggs. A few larvae (range 0–1) were detected in the eyes of both mice inoculated with B. potosis or B. transfuga eggs. The result indicated that B. potosis larvae do not show a higher tendency to migrate into the brain of mice than B. transfuga larvae. However, 1 mouse inoculated with 100 eggs had 1 larva in the central nervous system and showed a serious neurological sign. This result may underline a potential risk of B. potosis to cause neural larva migrans in humans.

Human babesiosis is a tick-borne protozoal disease of increasing clinical significance in North America. Most cases in the eastern and Midwestern regions of the United States are reportedly due to Babesia microti infections. By contrast, most human infections reported in California and Washington have been attributed to a new species that was first identified in 1991 and subsequently named Babesia duncani. Although the tick vector and mammalian reservoir hosts for B. microti are well characterized, the vector and reservoir hosts for B. duncani are unknown. As a result, specific risk factors for human infections cannot be characterized. Identification of potential hosts and vector species has been hampered by the lack of specific and sensitive molecular diagnostic tools to amplify parasite DNA. To address this need, a nested PCR assay targeting the β-tubulin gene, a well-conserved locus in piroplasm parasites with a highly variable intron region among species, was developed. The assay was evaluated by spiking tick and mammalian DNA extracts with DNA from a B. duncani isolate derived from a human patient (WA-1) as well as related Babesia spp. from Californian wildlife. This assay was highly specific, with a sensitivity of approximately 1 copy of template DNA in a background of tick DNA. At this level of detection B. duncani was detectable in larval tick samples, and the target locus allowed for visual differentiation between species by gel electrophoresis. This assay offers researchers a new tool for elucidating the natural transmission cycle of B. duncani.

The objective of this study was to report a case of a hydronephrotic patient with Strongyloides stercoralis infection, with discharge of rhabditoid larvae exclusively in urine. In 2013, a 72-yr-old male patient, hypertensive, obese, and diagnosed with hydronephrosis secondary to renal calculi, reported lumbar pain, polyuria, polaciuria, and dysuria, as well as frequent urinary tract infections. The microscopic analysis of urine sediment showed the presence of S. stercoralis rabditoid larvae. However, parasitological examinations by Baermann–Moraes, agar plate culture, and spontaneous sedimentation performed with 3 fecal samples on alternate days had negative results. The patient was treated with albendazole and to date has shown negative results in both parasitological and urine tests. This report deals with the unusual finding of S. stercoralis in a urine sample of an immunocompetent individual and absence of disseminated infection, but with hydronephrosis. Patients with nephropathies from S. stercoralis-endemic areas should be monitored periodically, as early detection may prevent the worsening of symptoms and renal failure.