Genetic Diversity of Clinical and Environmental Vibrio parahaemolyticus Strains from the Pacific Northwest

Since 1997, cases of Vibrio parahaemolyticus-related gastroenteritis from the consumption of raw oysters harvested in Washington State have been higher than historical levels. These cases have shown little or no correlation with concentrations of potentially pathogenic V. parahaemolyticus (positive for the thermostable direct hemolysin gene, tdh) in oysters, although significant concentrations of tdh⁺ V. parahaemolyticus strains were isolated from shellfish-growing areas in the Pacific Northwest (PNW). We compared clinical and environmental strains isolated from the PNW to those from other geographic regions within the United States and Asia for the presence of virulence-associated genes, including the thermostable direct hemolysin (tdh), the thermostable-related hemolysin (trh), urease (ureR), the pandemic group specific markers orf8 and toxRS, and genes encoding both type 3 secretion systems (T3SS1 and T3SS2). The majority of clinical strains from the PNW were positive for tdh, trh, and ureR genes, while a significant proportion of environmental isolates were tdh⁺ but trh negative. Hierarchical clustering grouped the majority of these clinical isolates into a cluster distinct from that including the pandemic strain RIMD2210633, clinical isolates from other geographical regions, and tdh⁺, trh-negative environmental isolates from the PNW. We detected T3SS2-related genes (T3SS2β) in environmental strains that were tdh and trh negative. The presence of significant concentrations of tdh⁺, trh-negative environmental strains in the PNW that have not been responsible for illness and T3SS2β in tdh- and trh-negative strains emphasizes the diversity in this species and the need to identify additional virulence markers for this bacterium to improve risk assessment tools for the detection of this pathogen.     

Palynomorphs from the Cambrian Series 3, Parahio valley (Spiti), Northwest Himalaya

We present low-diversity acritarch assemblages, including Alliumella baltica, Bavlinella faveolata, Brocholaminaria nigrita, Brocholaminaria sp., Eliasum sp., Leiosphaeridia minutissima, Leiosphaeridia spp., Lophosphaeridium sp., Pterospermella solida, Satka colonialica, Siphonophycus sp., and Synsphaeridium sp., as well as filamentous algae and cryptospore-like microfossils recorded from 45 rock samples around the Oryctocephalus indicus Zone (Cambrian Series 3) in the Cambrian Kunzam La (Parahio) Formation at the Kaltarbo locality in the Parahio valley, Northwest Himalaya. One new species Synsphaeridium parahioense was established. The acritarch assemblages at the basal part of the Cambrian Series 3 in the Kunzam La (Parahio) Formation were dependent on the local marine environment. The new discovery of cryptospore-like microfossils from the Cambrian Kunzam La (Parahio) Formation supports the extensive distribution of possible cryptospores in the Cambrian stratigraphic sequences.     

中国西北潜在蒸散时空演变特征及其定量化成因

潜在蒸散是区域干湿状况评价、作物需水量估算和水资源合理规划的关键因子。基于FAO推荐的Penman-Monteith公式和126个台站1961—2009年逐日气象观测资料估算西北干旱半干旱区的潜在蒸散量ET0,并对其空间分布特征进行了讨论。通过非参数化Sen趋势分析法和M-K统计检验法方法分析潜在蒸散的时间演变规律,并定量探讨了西北地区影响ET0变化的主导因素。结果表明:49 a来,西北地区ET0的年平均值约为980.63 mm,其中夏季的值最大,冬季的值最小。年平均ET0的大值区位于西北日照时数的高值中心,低值区主要位于海拔高,气温低的山区。西北地形和气候的多样性导致多年平均潜在蒸散的变化及其原因具有明显的时空差异。ET0的变化主要归因于风速和气温,而相对湿度和日照时数的作用较小。由于风速的负贡献超过气温的正贡献,导致年平均ET0整体上呈下降趋势。四季中,春冬两季的ET0缓慢上升,冬季的变化率是春季的两倍;夏秋两季的ET0有所下降,但只有夏季的变化趋势显著。春、夏、秋三季ET0变化的首要主导因子是风速,而冬季的首要主导因子是气温。空间上,西风带气候区ET0降低主要归因于风速的减小,陕南地区ET0下降主要归因于日照时数的减少,其它地区ET0上升的主要原因是气温的增加。     

Discovery of Steninae from Ningxia,Northwest China (Coleoptera,Staphylinidae)

A study on the Steninae of Ningxia Autonomous Region is presented. Sixteen species are recognized, including new province records for 11 species and four new species: Stenus biwenxuani sp. n., Stenus liupanshanus sp. n., Dianous yinziweii sp. n., Dianous ningxiaensis sp. n. Habitus photos of the new species, illustrations of diagnostic characters of all species and a key to species of the Steninae recorded from Ningxia are provided.     

Nematodes in Dryland Field Crops in the Semiarid Pacific Northwest United States

Soils and roots of field crops in low-rainfall regions of the Pacific Northwest were surveyed for populations of plantparasitic and non-plant-parasitic nematodes. Lesion nematodes (Pratylenchus species) were recovered from 123 of 130 non-irrigated and 18 of 18 irrigated fields. Pratylenchus neglectus was more prevalent than P. thornei, but mixed populations were common. Population densities in soil were affected by crop frequency and rotation but not by tillage or soil type (P < 0.05). Many fields (25%) cropped more frequently than 2 of 4 years had potentially damaging populations of lesion nematodes. Pratylenchus neglectus density in winter wheat roots was inversely correlated with grain yield (r² = 0.64, P = 0.002), providing the first field-derived evidence that Pratylenchus is economically important in Pacific Northwest dryland field crops. Stunt nematodes (Tylenchorhynchus clarus and Geocenamus brevidens) were detected in 35% of fields and were occasionally present in high numbers. Few fields were infested with pin (Paratylenchus species) and root-knot (Meloidogyne naasi and M. chitwoodi) nematodes. Nematodes detected previously but not during this survey included cereal cyst (Heterodera avenae), dagger (Xiphinema species), and root-gall (Subanguina radicicola) nematodes.     

Discovery of a Late Ordovician Foliomena fauna in the Tarim desert,Northwest China

A Foliomena fauna is reported for the first time from the Tarim paleoplate, and stratigraphically from the Yinpingshan Formation (upper Katian, Upper Ordovician) of Querqueke, Kuruktag region, northeastern Tarim, southern Xinjiang, Northwest China. The fauna includes seven species of brachiopods, amongst which three are new and four indeterminate: Anomaloglossa? sp., Orbiculoidea? sp., Foliomena xinjiangensis n. sp., Sericoidea minuta n. sp., Kassinella tarimensis n. sp., Rostricellula? sp., and Anazyga? sp. These species formed a Sericoidea-Kassinella Association, characterized by very small and well-preserved brachiopods, and well-developed laminations in its hosted mudstone, indicating a deep water environment (corresponding to lower BA5 to BA6). The faunal and sedimentological features suggest its affinity to the typical Foliomena faunal group of deep water origin. Numerical analyses show that the Foliomena fauna in late Katian time differentiated into two major paleogeographically related groups, and the Tarim association has a close faunal affinity to the representatives of this fauna in South China, indicating an active faunal exchange between Tarim and South China before the end-Ordovician mass extinction.     

Coronavirus and paramyxovirus in bats from Northwest Italy

Background

Bat-borne virus surveillance is necessary for determining inter-species transmission risks and is important due to the wide-range of bat species which may harbour potential pathogens. This study aimed to monitor coronaviruses (CoVs) and paramyxoviruses (PMVs) in bats roosting in northwest Italian regions. Our investigation was focused on CoVs and PMVs due to their proven ability to switch host and their zoonotic potential. Here we provide the phylogenetic characterization of the highly conserved polymerase gene fragments.

Results

Family-wide PCR screenings were used to test 302 bats belonging to 19 different bat species. Thirty-eight animals from 12 locations were confirmed as PCR positive, with an overall detection rate of 12.6% [95% CI: 9.3–16.8]. CoV RNA was found in 36 bats belonging to eight species, while PMV RNA in three Pipistrellus spp. Phylogenetic characterization have been obtained for 15 alpha- CoVs, 5 beta-CoVs and three PMVs; moreover one P. pipistrellus resulted co-infected with both CoV and PMV. A divergent alpha-CoV clade from Myotis nattereri SpA is also described. The compact cluster of beta-CoVs from R. ferrumequinum roosts expands the current viral sequence database, specifically for this species in Europe. To our knowledge this is the first report of CoVs in Plecotus auritus and M. oxygnathus, and of PMVs in P. kuhlii.

Conclusions

This study identified alpha and beta-CoVs in new bat species and in previously unsurveyed Italian regions. To our knowledge this represents the first and unique report of PMVs in Italy. The 23 new bat genetic sequences presented will expand the current molecular bat-borne virus databases. Considering the amount of novel bat-borne PMVs associated with the emergence of zoonotic infections in animals and humans in the last years, the definition of viral diversity within European bat species is needed. Performing surveillance studies within a specific geographic area can provide awareness of viral burden where bats roost in close proximity to spillover hosts, and form the basis for the appropriate control measures against potential threats for public health and optimal management of bats and their habitats.

     

Genetic Population Structure of Pacific Hake, Merluccius productus, in the Pacific Northwest

This study presents the first protein electrophoretic study of population structure within the Georgia Basin Pacific hake Distinct Population Segment, as defined under the U.S. Endangered Species Act. Forty-one allozyme loci (29 polymorphic) were analyzed in samples from three Pacific hake spawning populations on the west coast of North America: (1) Port Susan, Puget Sound, Washington (three temporal samples); (2) south-central Strait of Georgia, British Columbia, Canada (two temporal samples); and (3) offshore of southern California (two temporal samples) (total n = 664). Mean heterozygosity over all loci was 12–13% for all populations. Within-population temporal samples were not significantly different from one another, but statistically significant differences were detected at 15 of the 29 polymorphic loci (p < 0.05) among the three populations. Differences at eight of these loci were highly significant (p < 0.001): ADA ^*, ALAT ^*, bGALA ^*, GPI-A ^*, sIDHP ^*, LDH-A ^*, MPI ^*, and PEP-B ^*. The two Georgia Basin populations were significantly different at six loci: bGALA ^*, sIDHP ^*, LDH-A ^*, MPI ^*, PGK ^*, and PGM-2 ^* (p < 0.05). Nei's genetic distance (D) was 0.0006 between Port Susan and Strait of Georgia pooled temporal samples, and 0.005 between these populations and offshore Pacific hake. F_ST was 0.02 and 0.0046 among all three populations and among the Georgia Basin populations, respectively. Both F_ST estimates were significantly greater than zero, and the results suggest a high degree of demographic isolation among all three populations.     

Variation of 15N natural abundance in leaves and nodules of actinorhizal shrubs in Northwest Patagonia

This study was performed to assess the N₂-fixing capability of the native actinorhizal species Ochetophila trinervis (sin. Discaria trinervis) and Discaria chacaye (Rhamnaceae) in Northwest Patagonia. We measured the N concentration and ¹⁵N natural abundance in leaves and nodules of O. trinervis and D. chacaye, in leaves of associated non-actinorhizal vegetation, and in the soils under each sampled plant. O. trinervis and D. chacaye had foliar N concentrations that were about twice that of non-actinorhizal shrubs growing at the same sites, even though soils varied four-fold in total N across the sites. Leaves of both actinorhizal plants had a similar δ¹⁵N at any site and were close to atmospheric values. The foliar δ¹⁵N of non-actinorhizal plants and soil δ¹⁵N were strongly correlated across the sites. Nodules were depleted in δ¹⁵N relative to the foliage of the respective actinorhizal species. In conjunction with the uniformly high foliage N concentration of these actinorhizal plants and the universal presence of vesicles observed in root nodules, these data strongly suggest that O. trinervis and D. chacaye obtain a significant amount of their N from N₂ fixation. To calculate the proportion of N derived from atmosphere, theoretical B-values were estimated. In all cases where the δ¹⁵N of fixing and reference foliage were significantly different, O. trinervis and D. chacaye obtained almost all of their N from N₂ fixation. These results are the first to demonstrate N₂ fixation by O. trinervis and D. chacaye in the field and therefore suggest an important role for these actinorhizal plants in the N economy of ecosystems in northwest Patagonia as well as their potential use for restoration of degraded lands in this region.     

Early Carboniferous spiriferoid brachiopods from the Qaidam Basin,Northwest China: Taxonomy,biostratigraphy and biogeography

Six Early Carboniferous brachiopod species in four genera of the Superfamily Spiriferoidea are described from the Qaidam Basin, northwestern China, including a new genus, Qaidamospirifer, and two new species: Grandispirifer qaidamensis and Qaidamospirifer elongatus. Additionally, a new genus, Triangulospirifer, is also proposed to replace Triangularia Poletaev, 2001 that was preoccupied by a Devonian molluscan genus.On the basis of the new material as well as published information, we have reviewed the taxonomic composition and the stratigraphic and palaeobiogeographic distributions of the three previously established genera from the viewpoint of palaeobiogeography. The study reveals that Grandispirifer has a relatively long stratigraphic range from the late Tournaisian to Serpukhovian. During this interval, the genus attained a wide geographical distribution, reaching Northwest China, western Yunnan of West China, Japan, as well as Iran and North Africa. Angiospirifer first occurred in western Europe in the Viséan, and later migrated to North Africa during the late Viséan. In the Serpukhovian, it migrated eastward, reaching the Donets Basin of Ukraine and the Qaidam Basin in Northwest China. Anthracothyrina evolved from Brachythyrina in North Africa in late Viséan, then dispersed north-westward to western and eastern Europe and, further eastward to the Qaidam Basin during the Serpukhovian.     

Nutrient Limitation of Native and Invasive N2-Fixing Plants in Northwest Prairies

Nutrient rich conditions often promote plant invasions, yet additions of non-nitrogen (N) nutrients may provide a novel approach for conserving native symbiotic N-fixing plants in otherwise N-limited ecosystems. Lupinus oreganus is a threatened N-fixing plant endemic to prairies in western Oregon and southwest Washington (USA). We tested the effect of non-N fertilizers on the growth, reproduction, tissue N content, and stable isotope δ¹⁵N composition of Lupinus at three sites that differed in soil phosphorus (P) and N availability. We also examined changes in other Fabaceae (primarily Vicia sativa and V. hirsuta) and cover of all plant species. Variation in background soil P and N availability shaped patterns of nutrient limitation across sites. Where soil P and N were low, P additions increased Lupinus tissue N and altered foliar δ¹⁵N, suggesting P limitation of N fixation. Where soil P was low but N was high, P addition stimulated growth and reproduction in Lupinus. At a third site, with higher soil P, only micro- and macronutrient fertilization without N and P increased Lupinus growth and tissue N. Lupinus foliar δ¹⁵N averaged −0.010‰ across all treatments and varied little with tissue N, suggesting consistent use of fixed N. In contrast, foliar δ¹⁵N of Vicia spp. shifted towards 0‰ as tissue N increased, suggesting that conditions fostering N fixation may benefit these exotic species. Fertilization increased cover, N fixation, and tissue N of non-target, exotic Fabaceae, but overall plant community structure shifted at only one site, and only after the dominant Lupinus was excluded from analyses. Our finding that non-N fertilization increased the performance of Lupinus with few community effects suggests a potential strategy to aid populations of threatened legume species. The increase in exotic Fabaceae species that occurred with fertilization further suggests that monitoring and adaptive management should accompany any large scale applications.     

Complex genetic structures of Sardinella zunasi in the Northwest Pacific detected by AFLP markers

Seventy-one Sardinella zunasi individuals collected from 5 locations in the Northwest Pacific were used in an AFLP analysis to examine the population genetic structure of S. zunasi. Using 4 selective primer combinations, 414 putative loci were detected, including 340 polymorphic loci (82.13%). Both AMOVA and pairwise F_ST analysis detected significant population genetic structure between Chinese and Japanese populations, supporting separate stocks in this species. The UPGMA trees also detected significant spatial structure within S. zunasi. Pleistocene glaciations are hypothesized to be responsible for the genetic differentiation. The significant genetic differentiation found among S. zunasi populations might also be due to geographic isolation, because a pattern of isolation-by-distance was observed. The weaker but significant structures between the Japanese Aichi and Kagawa populations might be impacted by the semi-enclosed coastal topography there. No significant genetic structure was detected among the Chinese populations, perhaps because of S. zunasi’s migratory behavior and ocean current patterns.     

Genetic and phytochemical diversities of Cynomorium songaricum Rupr. in Northwest China indicated by ISSR markers and HPLC-fingerprinting

Cynomorium songaricum Rupr. is a parasitic plant and its most common host is Nitraria tangutorum Bobr. which grows in desert areas. To support the sustainable use and conservation of C. songaricum, genetic and phytochemical studies were undertaken on 16 different populations. Samples were analyzed using inter-simple sequence repeat (ISSR) markers and high performance liquid chromatography (HPLC)-fingerprinting. Total phenolic acids were determined by UV spectrophotometry. At the population level, percentage of polymorphic bands (PPB) ranged from 29.89 to 64.94%, Nei's genetic diversity index (H) ranged from 0.0931 to 0.2055, Shannon's information index (I) ranged from 0.1422 to 0.3140. 16 populations were classified into four chemotypes according to HPLC-fingerprinting analysis. Total phenolic acids contents ranged from 56.79 to 123.50 mg g⁻¹. The results of the genetic and chemical studies are used to suggest a planting and conservation system for C. songaricum.     

Prevalence of Blastocystis in Shelter-Resident and Client-Owned Companion Animals in the US Pacific Northwest

Domestic dogs and cats are commonly infected with a variety of protozoan enteric parasites, including Blastocystis spp. In addition, there is growing interest in Blastocystis as a potential enteric pathogen, and the possible role of domestic and in-contact animals as reservoirs for human infection. Domestic animals in shelter environments are commonly recognized to be at higher risk for carriage of enteropathogens. The purpose of this study was to determine the frequency of infection of shelter-resident and client-owned domestic dogs and cats with Blastocystis spp in the Pacific Northwest region of the USA. Fecal samples were collected from 103 shelter-resident dogs, 105 shelter-resident cats, 51 client-owned dogs and 52 client-owned cats. Blastocystis were detected and subtypes assigned using a nested PCR based on small subunit ribosomal DNA sequences. Shelter-resident animals were significantly more likely to test positive for Blastocystis (P<0.05 for dogs, P = 0.009 for cats). Sequence analysis indicated that shelter-resident animals were carrying a variety of Blastocystis subtypes. No relationship was seen between Blastocystis carriage and the presence of gastrointestinal disease signs in either dogs or cats. These data suggest that, as previously reported for other enteric pathogens, shelter-resident companion animals are a higher risk for carriage of Blastocystis spp. The lack of relationship between Blastocystis carriage and intestinal disease in shelter-resident animals suggests that this organism is unlikely to be a major enteric pathogen in these species.     

Molecular survey and characterization of Theileria annulata and Ehrlichia ruminantium in cattle from Northwest China

Theileriosis and ehrlichiosis are two important tick-borne diseases affecting cattle farming in China. However, limited information is available regarding prevalence and molecular characterization of Theileria annulata and Ehrlichia ruminantium in cattle in Xinjiang Uygur Autonomous Region (XUAR), northwestern China. In this study, a total of 176 blood samples of cattle from three rural areas of XUAR were collected in June 2017 and were tested by nested-PCR. A total of 34 (19.3%) samples were found to be infected with one or two pathogens. The overall prevalence rates of T. annulata and E. ruminantium were 18.2% and 1.7%, respectively. Phylogenetic analyses revealed that the E. ruminantium isolates from XUAR were located in the same clade but diverged from the isolates from African countries using pCS20 gene while T. annulata isolates from XUAR revealed differences in the genotypes using Tams1 sequences. To our knowledge, this is the first report of E. ruminantium infection in cattle in China. It also provides the first genetic characterization of T. annulata in cattle in XUAR. The current findings are important for understanding the distribution of agents of theileriosis and ehrlichiosis and in designing measures for the prevention and control of tick-borne diseases in cattle, other animals, and humans.     

Nitrite Reductase Genes (nirK and nirS) as Functional Markers To Investigate Diversity of Denitrifying Bacteria in Pacific Northwest Marine Sediment Communities

Genetic heterogeneity of denitrifying bacteria in sediment samples from Puget Sound and two sites on the Washington continental margin was studied by PCR approaches amplifying nirK and nirS genes. These structurally different but functionally equivalent single-copy genes coding for nitrite reductases, a key enzyme of the denitrification process, were used as a molecular marker for denitrifying bacteria. nirS sequences could be amplified from samples of both sampling sites, whereas nirK sequences were detected only in samples from the Washington margin. To assess the underlying nir gene structure, PCR products of both genes were cloned and screened by restriction fragment length polymorphism (RFLP). Rarefraction analysis revealed a high level of diversity especially for nirS clones from Puget Sound and a slightly lower level of diversity for nirK and nirS clones from the Washington margin. One group dominated within nirK clones, but no dominance and only a few redundant clones were seen between sediment samples for nirS clones in both habitats. Hybridization and sequencing confirmed that all but one of the 228 putative nirS clones were nirS with levels of nucleotide identities as low as 45.3%. Phylogenetic analysis grouped nirS clones into three distinct subclusters within the nirS gene tree which corresponded to the two habitats from which they were obtained. These sequences had little relationship to any strain with known nirS sequences or to isolates (mostly close relatives of Pseudomonas stutzeri) from the Washington margin sediment samples. nirK clones were more closely related to each other than were the nirS clones, with 78.6% and higher nucleotide identities; clones showing only weak hybridization signals were not related to known nirK sequences. All nirK clones were also grouped into a distinct cluster which could not be placed with any strain with known nirK sequences. These findings show a very high diversity of nir sequences within small samples and that these novel nir clusters, some very divergent from known sequences, are not known in cultivated denitrifiers.     

Molecular Phylogeography and Evolutionary History of the Estuarine Copepod, Acartia Tonsa, on the Northwest Atlantic Coast

Coastal estuaries are useful model systems to study the ecological and evolutionary responses of organisms to highly variable, discontinuous habitats. For this study, the molecular population genetic diversity of the planktonic calanoid copepod Acartia tonsa (Dana, 1849) was described based on DNA sequence variation for a 183 base-pair region of the mitochondrial 16S rRNA gene. Samples of A. tonsa were collected from four estuaries on the Atlantic coast of the USA during 1993 and 1994, one estuary on the Gulf of Mexico coast in 1994, and one site on the Pacific coast of the USA in 1994. Dispersal of A. tonsa was shown to be restricted, with significant population genetic structuring between different estuaries. For all but the closely-adjacent MA and RI samples, frequencies of haplotypes and/or length polymorphisms within one haplotype (caused by insertion/deletion mutations) revealed highly significant genetic differentiation and geographic isolation. Mt16S haplotypes of A. tonsa from Atlantic and Gulf of Mexico estuaries were assorted among four deeply-diverged clades. Haplotypes within each clade differed by <2%, while differences among clades of 10% to 14% approached those between described Acartia species (e.g., 19% to 28% among A. clausi, A. hudsonica, and A. longiremis). Atlantic and Pacific coast samples identified as A. tonsa had no haplotypes in common and genetic differences between haplotypes ranged from 18% to 29%; phylogenetic analysis supported the separation of Pacific coast A. tonsa as a distinct species. We hypothesize that the observed patterns of molecular genetic diversity and structure of A. tonsa resulted from responses to historical climatic variation, including episodic range compression and displacement, and alteration of NW Atlantic coastal and estuarine environments.     

Age of the Silurian Wuxiahe Formation in Langao,Northwest China: New conodont data

A recent study of conodonts from the Wuxiahe Formation (lower to middle part) in the Ziyang-Langao region suggested its age of middle Telychian (Llandovery) to lower Sheinwoodian (Wenlock), contradicted by subsequent graptolite studies indicating an age of late Telychian for the same interval. New samples from the Qiaoxi section for conodonts to re-access the age of the Wuxiahe Formation collected in this study show that the lower to middle part of the formation belongs to the Pterospathodus amorphognathoides amorphognathoides Biozone, suggesting the age of late Telychian; thus, the Llandovery–Wenlock boundary in the section is most probably higher than previously estimated, but its precise position is not determined since the identification of the Wenlock graptolite Cyrtograptus cf. lundgreni in the section is to be further confirmed. Based on the conodont faunas recognized in the Qiaoxi and Tianwancun sections, the base of the Wuxiahe Formation in the Ziyang-Langao region is diachronous, i.e., not higher than the upper Telychian Pterospathodus amorphognathoides amorphognathoides Biozone at Qiaoxi, but not lower than the lower Sheinwoodian Kockelella ranuliformis Biozone at the Tianwancun setion.     

Chagas Disease in 2 Geriatric Rhesus Macaques (Macaca mulatta) Housed in the Pacific Northwest

Chagas disease (American trypanosomiasis) is caused by the protozoan parasite Trypanosoma cruzi. It is endemic in Latin America but also is found in the southern United States, particularly Texas and along the Gulf Coast. Typical clinical manifestations of Chagas disease are not well-characterized in rhesus macaques, but conduction abnormalities, myocarditis, and encephalitis and megaesophagus have been described. Here we report 2 cases of Chagas disease in rhesus macaques housed in the northwestern United States. The first case involved a geriatric male macaque with cardiomegaly, diagnosed as dilated cardiomyopathy on ultrasonographic examination. Postmortem findings included myocarditis as well as ganglioneuritis in the esophagus, stomach, and colon. The second case affected a geriatric female macaque experimentally infected with SIV. She was euthanized for a protocol-related time point. Microscopic examination revealed chronic myocarditis with amastigotes present in the cardiomyocytes, ganglioneuritis, and opportunistic infections attributed to her immunocompromised status. Banked serum samples from both macaques had positive titers for T. cruzi. T. cruzi DNA was amplified by conventional PCR from multiple tissues from both animals. Review of their histories revealed that both animals had been obtained from facilities in South Texas more than 12 y earlier. Given the long period of clinical latency, Chagas disease may be more prevalent in rhesus macaques than typically has been reported. T. cruzi infection should be considered for animals with unexplained cardiac or gastrointestinal pathology and that originated from areas known to have a high risk for disease transmission.Abbreviations: DCM, dilated cardiomyopathy; CMV, cytomegalovirus; NHP, nonhuman primateChagas disease is caused by the hemoflagellate protozoan parasite Trypanosoma cruzi. The disease is endemic in many regions of South and Central America, and its range extends to the southern United States. In the United States, there is evidence that the parasite has established a domestic transmission cycle with canine reservoirs,¹⁹ and there are numerous wildlife reservoirs, most importantly armadillos, raccoons, rodents, and opossums.⁶ The main mode of transmission is via arthropod vectors, primarily triatomine species (kissing bugs or cone-nosed bugs), which serve as intermediate hosts. Vector species are present in the southern half of the United States.² Infection has been reported sporadically in domestic nonhuman primate (NHP) colonies.¹² Autochthonous insect vector-mediated transmission in humans in the United States has been reported rarely.³³ Transmission of T. cruzi to NHP is thought to occur mainly through insect vectors, specifically by contamination of the oral mucous membranes with parasite-containing feces during consumption of the bug. The infection may remain subclinical for years and, similar to that in people, affects the nervous system, digestive system, and heart. Clinical findings in NHP are infrequent but can include subcutaneous edema, fever, anorexia, lethargy, heart failure, and sudden death.^4,5 As in humans, the disease in NHP consists of an acute phase, with a paucity of clinical manifestations, and a chronic phase, characterized by irreversible cardiomyopathy leading to cardiac dysfunction and death. Chronically infected NHP in the indeterminate form of the chronic phase can exhibit subclinical conduction and echocardiographic abnormalities.⁸T. cruzi infections have been reported in a number of NHP species housed in Texas, Louisiana, and Georgia. Species affected include rhesus macaques (Macaca mulatta),^12,17 cynomolgus macaques (M. fascicularis),^29,42 yellow baboons (Papio cynocephalus), olive baboons (P. anubis),^12,14,41 pig-tailed macaques (M. nemestrina),^12,35 squirrel monkeys (Saimiri sciureus),¹³ ring-tailed lemurs (Lemur catta),^15,30 black-eyed lemurs (Eulemur macaco flavifrons), black and white ruffed lemurs (Varecia variegata variegate),¹⁵ pileated gibbon (Hylobates pileatus),³⁶ a lion-tailed macaque (M. silenus),³⁰ a Celebes crested black macaque (M. nigra),²⁷ and a chimpanzee (Pan troglodytes).⁴ Here we report on 2 cases of Chagas disease in rhesus macaques housed in the northwestern United States but that originated from South Texas.     

Cytogeography and phenomenon of cytomixis in Silene vulgaris from cold regions of Northwest Himalayas (India)

Silene vulgaris is an important ethnobotanical species, but so far no thorough meiotic analysis has been performed, despite its wide occurrence in temperate and alpine Himalayas, this prompted us to undertake present study. We investigated original meiotic chromosome number and effect of cytomixis on meiotic course, microsporogenesis and pollen grain size and fertility. The paper also summarizes the previously published chromosome counts in the species and cytogeography of 2x and 4x cytotypes in the Indian Himalayas and elsewhere. Meiocytes preparations were made through standard squashed technique in 1 % acetocarmine. Two ploidy levels have been determined during the cytomorphological surveys, diploid (2n = 2x = 24) and tetraploid (2n = 4x = 48). Most of the presently scored populations were found to be diploid (2n = 2x = 24). Morphologically 2x and 4x individuals were indistinguishable from each other. Meiosis was perfectly normal in 4x cytotype. However, some of the 2x plants showed the phenomenon of cytomixis and associated meiotic abnormalities. Consequent to cytomixis, hypo-, hyperploid and enucleated pollen mother cells (PMCs) were resulted. Such PMCs combined with other meiotic abnormalities yielded sterile and variable sized pollen grains. On the basis of overall information, it is clear that 2x cytotype is the most widely distributed cytotype in the Indian Himalayas as well as elsewhere. On the other hand, the 4x cytotype seems to be less frequently present in the Indian Himalayas and outside of India.