Cost-effectiveness analysis of arterial catheter insertion on robotic-assisted laparoscopic prostatectomy.

INTRODUCTION: To evaluate the utility, outcomes, and cost of arterial line placement in a single institution cohort of patients undergoing robotic-assisted laparoscopic prostatectomy (RALP). MATERIALS AND METHODS: A retrospective chart review was performed at a large tertiary care center from July 2018 through January 2021. Hospital costs and cost-effective analysis was performed on patients with and without arterial line placement. Means with standard deviations were used to report continuous variables, while numbers and percentages were utilized to describe categorical variables. T-tests and Chi-square tests compared categorical and continuous variables across study cohorts, respectively. Multivariable analyses were used to examine the association between A-line placement and outcomes as mentioned above adjusting for the effect of other co-variables. RESULTS: Among the 296 included patients, 138 (46.6%) had arterial lines. No preoperative patient characteristic predicted arterial line placement. Rates of complications and re-admissions were not statistically significant between the two groups. Arterial line use was associated with higher volumes of intraoperative fluid administration, as well as a longer hospital length of stay. Total cost and operative time did not significantly differ between cohorts, but arterial line placement increased variability of these factors. CONCLUSION: The use of arterial lines in patients undergoing RALP is not necessarily guideline-driven and does not decrease the rate of perioperative complications. However, it is associated with longer length of stay and increases variability in charge. These data show that the surgical team and anesthesia team should critically evaluate the need for arterial line placement in patients undergoing RALP.

Clinical and Immunologic Outcomes After Immediate or Deferred Antiretroviral Therapy Initiation During Primary Human Immunodeficiency Virus Infection: The Sabes Randomized Clinical Study.

BACKGROUND: In addition to demonstrated public health benefits on reducing transmission, it remains unclear how early antiretroviral therapy (ART) must be started after acquisition of human immunodeficiency virus (HIV) to maximize individual benefits. METHODS: We conducted an open-label randomized clinical study in Lima, Peru among adult men who have sex with men and transgender women with acute (HIV-antibody negative/HIV-1 RNA positive) or recent (confirmed negative HIV-antibody or RNA test within 3 months) HIV infection, who were randomized to start ART immediately versus defer by 24 weeks. We evaluated outcomes by treatment arm and immunologic markers by days since estimated date of detectible infection (EDDI). RESULTS: Of 216 participants, 105 were assigned to immediate arm and 111 to deferred arm (median age 26.8 years, 37% with acute HIV). The incidence of non-ART-related adverse events was lower in immediate versus deferred arm (83 vs 123/100 person-years, IRR 0.67 (95% confidence interval [CI] .47, .95; P = .02), the difference dominated by fewer infections in those treated immediately. After 24 weeks of ART, between-group differences in CD4/CD8 cell ratio lessened (P = .09 overall), but differences between those initiating ART ≤ 30 days from EDDI (median 1.03, interquartile range [IQR] 0.84, 1.37), and those initiating > 90 days (0.88, IQR 0.61, 1.11) remained, P = .02. Principal components analysis of 20 immune biomarkers demonstrated distinct patterns between those starting ART > 90 days from EDDI versus those starting within 30 or 90 days (both P < .001). CONCLUSIONS: To our knowledge, this is the only evaluation of randomized ART initiation during primary HIV and provides evidence to explicitly consider acute HIV in World Health Organization recommendations for universal ART. CLINICAL TRIALS REGISTRATION: NCT01815580.

Mechanisms of Endogenous HIV-1 Reactivation by Endocervical Epithelial Cells.

Pharmacological HIV-1 reactivation to reverse latent infection has been extensively studied. However, HIV-1 reactivation also occurs naturally, as evidenced by occasional low-level viremia ("viral blips") during antiretroviral treatment (ART). Clarifying where blips originate from and how they happen could provide clues to stimulate latency reversal more effectively and safely or to prevent viral rebound following ART cessation. We studied HIV-1 reactivation in the female genital tract, a dynamic anatomical target for HIV-1 infection throughout all disease stages. We found that primary endocervical epithelial cells from several women reactivated HIV-1 from latently infected T cells. The endocervical cells' HIV-1 reactivation capacity further increased upon Toll-like receptor 3 stimulation with poly(I·C) double-stranded RNA or infection with herpes simplex virus 2 (HSV-2). Notably, acyclovir did not eliminate HSV-2-induced HIV-1 reactivation. While endocervical epithelial cells secreted large amounts of several cytokines and chemokines, especially tumor necrosis factor alpha (TNF-α), CCL3, CCL4, and CCL20, their HIV-1 reactivation capacity was almost completely blocked by TNF-α neutralization alone. Thus, immunosurveillance activities by columnar epithelial cells in the endocervix can cause endogenous HIV-1 reactivation, which may contribute to viral blips during ART or rebound following ART interruption.IMPORTANCE A reason that there is no universal cure for HIV-1 is that the virus can hide in the genome of infected cells in the form of latent proviral DNA. This hidden provirus is protected from antiviral drugs until it eventually reactivates to produce new virions. It is not well understood where in the body or how this reactivation occurs. We studied HIV-1 reactivation in the female genital tract, which is often the portal of HIV-1 entry and which remains a site of infection throughout the disease. We found that the columnar epithelial cells lining the endocervix, the lower part of the uterus, are particularly effective in reactivating HIV-1 from infected T cells. This activity was enhanced by certain microbial stimuli, including herpes simplex virus 2, and blocked by antibodies against the inflammatory cytokine TNF-α. Avoiding HIV-1 reactivation could be important for maintaining a functional HIV-1 cure when antiviral therapy is stopped.

Influence of 5-Alpha Reductase Inhibitors on Prostate Cancer Detection with Magnetic Resonance Imaging: A Matched Cohort Study.

PURPOSE: We evaluated the influence of 5-alpha reductase inhibitors (5-ARIs) on the performance of magnetic resonance imaging (MRI) for detection of Gleason grade group (GG) ≥2 prostate cancer, and on apparent diffusion coefficient (ADC) maps. MATERIALS AND METHODS: This single center, retrospective study included men who had MRI for initial detection or active surveillance of prostate cancer. The study group included 59 men who used for 5-ARIs for ≥12 months, and the control group included 59 men who were matched for both MRI indication and biopsy results. DeLong's test was used to compare the area under the receiver operating characteristic curve (AUC) for detection of GG ≥2 cancer between the groups. Wilcoxon rank sum test was used for comparison of lesions apparent diffusion coefficient (ADC) metrics between the groups. RESULTS: MRI accuracy in the study group (AUC=0.778) was not significantly different compared to the control group (AUC=0.821; 95% CI for difference 0.22-0.13; p=0.636). In the control group, all ADC metrics were lower in lesions with GG ≥2 cancer on biopsy than in those with GG 1 cancer or negative results (p=0.001-0.01). In the study group, this difference was significant only when the mean ADC of the lesions was normalized by the ADC of urine (p=0.044). CONCLUSIONS: Long-term exposure to 5-ARIs does not seem to impair the detection of significant cancer on MRI but may affect the ability of ADC metrics to discriminate between lesions that harbor significant cancer and those that harbor insignificant cancer or benign tissue.

High genetic diversity of the entomopathogenic fungus Beauveria bassiana in Colima, Mexico.

The entomopathogenic fungus Beauveria bassiana is used widely as a biological control agent against a wide range of insect pests globally. In this study, 44 Beauveria isolates from the state of Colima, Mexico harbored in the "Colección de Hongos Entomopatógenos" of the "Centro Nacional de Referencia de Control Biológico" and from different substrates, insect-hosts, and localities were characterized with molecular markers. All isolates were identified using a Bayesian phylogenetic analysis of translation elongation factor 1-α (TEF) and nuclear intergenic Bloc region. Forty-three isolates were identified as B. bassiana and grouped into two sub-clades, i.e., AFNEO_1 (n = 22; previously defined as a clade with African and Neotropical origin) and Bb clade (n = 21; closely associated with ex-type strain ARSEF 1564), and one isolate was identified as B. pseudobassiana. The fixation index (FST = 0.493) established the genetic differentiation between AFNEO_1 and Bb clades. High genotype richness and genetic diversity of AFNEO_1 and Bb clades were revealed in sequence analysis of Bloc region and SSR genotyping. Moreover, the AFNEO_1 and Bb clades were confirmed as two independent clonally structured assemblages. Finally, the AMOVA detected no significant association between any combination of substrate, insect-host or geographical origin. High genetic variation of B. bassiana in Colima, Mexico could suggest a functional diversity among isolates that may include those effective against a specific insect pest.

First description of adults of the type species of the genus Glossocercus Chandler, 1935 (Cestoda: Gryporhynchidae).

The type species of the genus Glossocercus Chandler, 1935, G. cyprinodontis Chandler, 1935, was described as metacestode (larval stage) from the mesentery of the sheepshead minnow fish (Cyprinodon variegatus Lacépède) from Galveston Bay, Texas. The description was based on the morphology of the rostellar hooks; however, the features of the internal morphology of the proglottides could no be provided. In the present study we describe for the first time the features of the adult G. cyprinodontis from the intestine of Pelecanus occidentalis Linnaeus, Nycticorax nycticorax Linnaeus and Egretta rufescens Gmelin in Mexico. Glossocercus cyprinodontis possesses similar strobilar morphology with the two other congeneric species, both distributed in the Neartic and Neotropical regions, i.e. Glossocercus caribaensis (Rysavy et Macko, 1971) and Glossocercus auritus (Rudolphi, 1819). However, G. cyprinodontis differs mainly in the shape of the rostellar hooks (those of G. cyprinodontis possess the handle and the guard strongly sclerified compared to those of G. auritus and G. caribaensis) and their size (total length of 175-203 microm in G. cyprinodontis compared to 189-211 microm in G. caribaensis and 220-285 microm in G. auritus). Generic diagnosis of Glossocercus is emended: rostellar hooks in two rows with ten hooks of different shape and length in each, scolex large and globular, proglottides craspedote, wider than long, genital pores irregularly alternating, vagina transverse, surrounded by epithelial cells, ventral to cirrus-sac, uterus bar-shaped in mature proglottides, occupies all space between osmoregulatory ducts with eggs in gravid proglottides, ovary lobed in middle of proglottis, cirrus-sac elongate, between osmoregulatory canals, cirrus armed with spinitriches and apical tuft of slender spinitriches.

A NEW SPECIES OF CHIGGER MITE AND THE RECORD OF A GASTROINTESTINAL NEMATODE IN SCELOPORUS CAUTUS (SQUAMATA: PHRYNOSOMATIDAE) FROM THE CHIHUAHUAN DESERT PROVINCE IN COAHUILA, MEXICO.

The chigger species Acomatacarus coahuilensis n. sp. (Acariformes: Trombidiformes: Leeuwenhoekiidae) inhabiting the neck scales of the wary spiny lizard Sceloporus cautus Smith, 1938 (Squamata: Iguania: Phrynosomatidae), from Chihuahuan Desert Province, Coahuila, Mexico, is described herein. The new mite species is assigned to the genus AcomatacarusEwing, 1942, based on a palptarsus with 6 branched setae plus the tarsala (ω). This new species can be separated from the other species in this genus mainly based on the combination of the following characters: presence of mastisetae (MTa) on tarsus III and tibiala (φ) on tibia III, subterminala (ζ1), parasubterminala (z), and pretarsala (ζ2) on tarsus I, 2 genualae (σ1-2) on genu I, onychotriches and claw-like empodium of legs I-III, and absence of mastisetae on tibia III. In addition to the new species, the gastrointestinal nematode Skrjabinoptera phrynosoma (Ortlepp, 1922) Schulz, 1927 (Physalopteridae), was also found, which represents a new host record in the wary spiny lizard. This study reports the third species of the genus Acomatacarus recorded from Mexico and the first record of chigger mites and helminth parasites associated with Sc. cautus.

Acute retroviral syndrome is associated with lower CD4 + T cell nadir and delayed viral suppression, which are blunted by immediate antiretroviral therapy initiation.

OBJECTIVES: To describe the prevalence of acute retroviral syndrome (ARS) and associated findings during primary HIV, and explore the relationship of ARS to clinical, virological, and immunological outcomes within a longitudinal screen, retest and treat study that minimized ascertainment bias. DESIGN: We evaluated ARS symptoms and signs among 216 persons with acute and early incident HIV within the Sabes study of timing of antiretroviral therapy (ART) initiation during primary HIV in Peru. METHODS: We evaluated patient reported symptoms and signs during primary HIV and used logistic regression and generalized linear models to evaluate associations with CD4 + and CD8 + T cell counts, HIV viral load, and a panel of 23 soluble markers of immune activation. RESULTS: Sixty-one percent of participants had at least one ARS finding and 35% had at least 3. More ARS findings were reported in those enrolled within a month of estimated date of detectable infection (EDDI). Having more ARS signs/symptoms was associated with increased risk of CD4 + cell decrease below 350 cells/ml within the first 24 weeks, failure to suppress HIV viral load, and was most strongly associated with elevated IP-10. Immediate ART blunted effects on symptoms, CD4 + cell count and viral load, as associations were strongest in the arm that started ART after 24 weeks. Detrimental associations of ARS with CD4 + counts, and CD4 + /CD8 + ratio were not maintained at 2 or 4 years. CONCLUSIONS: ARS has marked associations with short-term immunologic function and virologic suppression, which were mitigated in participants randomized to initiate ART immediately during primary infection.

Multi-locus phylogeny of the catfish genus Ictalurus Rafinesque, 1820 (Actinopterygii, Siluriformes) and its systematic and evolutionary implications.

BACKGROUND: Ictalurus is one of the most representative groups of North American freshwater fishes. Although this group has a well-studied fossil record and has been the subject of several morphological and molecular phylogenetic studies, incomplete taxonomic sampling and insufficient taxonomic studies have produced a rather complex classification, along with intricate patterns of evolutionary history in the genus that are considered unresolved and remain under debate. RESULTS: Based on four loci and the most comprehensive taxonomic sampling analyzed to date, including currently recognized species, previously synonymized species, undescribed taxa, and poorly studied populations, this study produced a resolved phylogenetic framework that provided plausible species delimitation and an evolutionary time framework for the genus Ictalurus. CONCLUSIONS: Our phylogenetic hypothesis revealed that Ictalurus comprises at least 13 evolutionary units, partially corroborating the current classification and identifying populations that emerge as putative undescribed taxa. The divergence times of the species indicate that the diversification of Ictalurus dates to the early Oligocene, confirming its status as one of the oldest genera within the family Ictaluridae.

Dynamic immune markers predict HIV acquisition and augment associations with sociobehavioral factors for HIV exposure.

Prior studies attempting to link biomarkers of immune activation with risk of acquiring HIV have relied on cross sectional samples, most without proximity to HIV acquisition. We created a nested case-control study within the Sabes study in Peru, and assessed a panel of plasma immune biomarkers at enrollment and longitudinally, including within a month of diagnosis of primary HIV or matched timepoint in controls. We used machine learning to select biomarkers and sociobehavioral covariates predictive of HIV acquisition. Most biomarkers were indistinguishable between cases and controls one month before HIV diagnosis. However, levels differed between cases and controls at study entry, months to years earlier. Dynamic changes in IL-2, IL-7, IL-10, IP-10 and IL-12, rather than absolute levels, jointly predicted HIV risk when added to traditional risk factors, and there was modest effect modification of biomarkers on association between sociobehavioral risk factors and HIV acquisition.

Immune Activation in Primary Human Immunodeficiency Virus: Influence of Duration of Infection, Treatment, and Substance Use.

Background: Primary human immunodeficiency virus (HIV) is characterized by dynamic changes in viral load and innate and adaptive immune responses; it is unclear the extent to which time from acquisition to antiretroviral therapy (ART) initiation and substance use impact these immunologic changes. Methods: We studied plasma immune activation biomarkers, viral load, and CD4+ and CD8+ cell counts in participants from the Sabes primary infection study in Peru, who had been randomized to begin ART immediately after diagnosis vs 24 weeks later. We modeled influence of substance use and duration of HIV infection on biomarkers at baseline and over 24 weeks. Results: Compared to participants enrolled >30 days after HIV acquisition, participants enrolled during acute infection (≤30 days) had higher mean interferon (IFN)-γ and IFN-α2a (1.7-fold and 3.8-fold interquartile range [IQR] higher, respectively). Participants enrolled >30 days after HIV acquisition had higher mean baseline CD8+ cell count (2.7 times the IQR). Alcohol use (positive phosphatidylethanol level) was associated with elevated IFN-γ, tumor necrosis factor alpha (TNF-α), and interleukin 12p70 (IL-12p70), and smoking was associated with higher macrophage inflammatory protein 1α, TNF-α, and IL-12p70. Most biomarkers declined more quickly in participants who initiated ART immediately; however, substance use and duration of HIV infection at enrollment had little influence on rate of decline. Conclusions: IFN-γ and other biomarkers are elevated during early primary infection, when exposure to HIV antigens is high. Immune activation decreased most quickly in those who started ART during acute/early primary infection. Higher CD8+ cell counts and a trend toward higher soluble CD163 levels during the 30 days after acquisition suggest the onset of compensatory responses and immune exhaustion.

HIV Disease Dynamics and Markers of Inflammation and CNS Injury During Primary HIV Infection and Their Relationship to Cognitive Performance.

INTRODUCTION: Early systemic and central nervous system viral replication and inflammation may affect brain integrity in people with HIV, leading to chronic cognitive symptoms not fully reversed by antiretroviral therapy (ART). This study examined associations between cognitive performance and markers of CNS injury associated with acute HIV infection and ART. METHODS: HIV-infected MSM and transgender women (average age: 27 years and education: 13 years) enrolled within 100 days from the estimated date of detectable infection (EDDI). A cognitive performance (NP) protocol was administered at enrollment (before ART initiation) and every 24 weeks until week 192. An overall index of cognitive performance (NPZ) was created using local normative data. Blood (n = 87) and cerebrospinal fluid (CSF; n = 29) biomarkers of inflammation and neuronal injury were examined before ART initiation. Regression analyses assessed relationships between time since EDDI, pre-ART biomarkers, and NPZ. RESULTS: Adjusting for multiple comparisons, shorter time since EDDI was associated with higher pre-ART VL and multiple biomarkers in plasma and CSF. NPZ scores were within the normative range at baseline (NPZ = 0.52) and at each follow-up visit, with a modest increase through week 192. Plasma or CSF biomarkers were not correlated with NP scores at baseline or after ART. CONCLUSIONS: Biomarkers of CNS inflammation, immune activation, and neuronal injury peak early and then decline during acute HIV infection, confirming and extending results of other studies. Neither plasma nor CSF biomarkers during acute infection corresponded to NP scores before or after sustained ART in this cohort with few psychosocial risk factors for cognitive impairment.

Buprenorphine Increases HIV-1 Infection In Vitro but Does Not Reactivate HIV-1 from Latency.

BACKGROUND: medication-assisted treatment (MAT) with buprenorphine is now widely prescribed to treat addiction to heroin and other illicit opioids. There is some evidence that illicit opioids enhance HIV-1 replication and accelerate AIDS pathogenesis, but the effect of buprenorphine is unknown. METHODS: we obtained peripheral blood mononuclear cells (PBMCs) from healthy volunteers and cultured them in the presence of morphine, buprenorphine, or methadone. We infected the cells with a replication-competent CCR5-tropic HIV-1 reporter virus encoding a secreted nanoluciferase gene, and measured infection by luciferase activity in the supernatants over time. We also surveyed opioid receptor expression in PBMC, genital epithelial cells and other leukocytes by qPCR and western blotting. Reactivation from latency was assessed in J-Lat 11.1 and U1 cell lines. RESULTS: we did not detect expression of classical opioid receptors in leukocytes, but did find nociception/orphanin FQ receptor (NOP) expression in blood and vaginal lymphocytes as well as genital epithelial cells. In PBMCs, we found that at physiological doses, morphine, and methadone had a variable or no effect on HIV infection, but buprenorphine treatment significantly increased HIV-1 infectivity (median: 8.797-fold increase with 20 nM buprenorphine, eight experiments, range: 3.570-691.9, p = 0.0078). Using latently infected cell lines, we did not detect reactivation of latent HIV following treatment with any of the opioid drugs. CONCLUSIONS: our results suggest that buprenorphine, in contrast to morphine or methadone, increases the in vitro susceptibility of leukocytes to HIV-1 infection but has no effect on in vitro HIV reactivation. These findings contribute to our understanding how opioids, including those used for MAT, affect HIV infection and reactivation, and can help to inform the choice of MAT for people living with HIV or who are at risk of HIV infection.

Genetic diversification of acanthocephalans of the genus Floridosentis Ward 1953 (Acanthocephala: Neoechinorhynchidae), parasites of mullets from the Americas.

Adult worms of the genus Floridosentis are endoparasites of marine fishes of the genus Mugil and are broadly distributed in the Americas. Currently, Floridosentis includes two species, F. mugilis, distributed in the Gulf of Mexico and along the Atlantic Ocean coast, and F. pacifica, restricted to the Pacific Ocean coast. The aim of this study was to explore the species limit of both species of the genus Floridosentis, collected in 37 localities in eight countries: Mexico, Guatemala, El Salvador, Honduras, Nicaragua, Costa Rica, Ecuador and Venezuela. We sequenced 253 specimens to build a comprehensive dataset for three genes: the cytochrome c oxidase subunit I (cox 1) from mitochondrial DNA, the internal transcribed spacers ITS1 and ITS2 including the 5.8S gene (ITS region), and the D2 + D3 domains of the large subunit (LSU) of nuclear DNA. Maximum likelihood and Bayesian analyses with the cox 1 and concatenated (cox 1 + ITS+LSU) datasets were conducted. Two species delimitation methods were implemented, the Automatic Barcode Gap Discovery (ABGD), and Bayesian species delimitation (BPP), plus a haplotype network inferred with 253 specimens, allowing us to validate two nominal species of Floridosentis., F. mugilis, plus one linage distributed in the Gulf of Mexico and along the Atlantic Ocean coast, and F. pacifica, plus two additional lineages distributed along the Pacific Ocean coast. All these lineages are shared by both species of mullet (Mugil curema and M. cephalus). The currents in the Atlantic Ocean, Pacific Ocean and Gulf of Mexico, in combination with the biology of the definitive hosts, have played a key role in the distribution of the two nominal species and of the three lineages of Floridosentis across the Americas.

A New Species of Morelacarus (Acariformes: Prostigmata: Leeuwenhoekiidae) Associated With Sceloporus grammicus (Squamata: Phrynosomatidae) From the Mexican Plateau, Zacatecas, Mexico.

A new species of chigger mites, Morelacarus uazi n. sp., parasitizing lizards of the species Sceloporus grammicus from Mazapil, Zacatecas, Mexico, is described and illustrated. This new taxon differs from other species of the genus by having a palpal claw with 4 subequal paired prongs, and it can be further distinguished by the combination of the following characters: presence of onychotriches and mastitarsala III, absence of mastitibiala III, subterminala I, and parasubterminala I, and the palpal setal formula B/B/BBb. Additionally, an identification key for the species of the genus Morelacarus is presented. The new species represents the seventh species assigned to the genus and the first species described from a Mexican lizard.

Identification of leopard frogs (Anura: Ranidae: Lithobates) distributed in some localities of the Southern Mexican Plateau using mitochondrial DNA sequences.

Species of the genus Lithobates are difficult to identify, especially on the 'Rana pipiens' complex, because the morphological differences among the species are often subtle. In this study, we used 12S ribosomal RNA gene partial sequences to identify specimens of leopard frogs from some new localities (not sampled on previous studies) of the Southern Mexican Plateau, to carry out a phylogenetic analysis, and also a morphometric analysis of some morphological features were analyzed to evaluate their morphological variation. A phylogenetic analysis using partial sequences of 12S rDNA mitochondrial gene for 31 samples was carried out using Bayesian Inference, Maximum Parsimony, and Maximum Likelihood. In addition, 20 morphological lineal measurements from 97 specimens were analyzed by principal component analysis (PCA) to evaluate if the frogs studied are conspecific. Partial sequences of the 12S rDNA obtained from frogs distributed in the Southern Mexican Plateau show two haplotypes with a divergence of 0.4%. Phylogenetic hypothesis shows an exclusive group with a previous sequence of Lithobates montezumae. The PCA indicates that variables are not linearly correlated and specimens belong to a single group. Evidence found, let us consider that specimens studied belong to one species of the Lithobates montezumae subgroup, distributed principally in the Southern Mexican Plateau.

Genetic diversity of the Metarhizium anisopliae complex in Colima, Mexico, using microsatellites.

Metarhizium anisopliae is a complex of cryptic species with wide geographical distribution and versatile lifestyles. In this study, 45 isolates of the Metarhizium genus harbored in the "Colección de Hongos Entomopatógenos" of the "Centro Nacional de Referencia de Control Biológico" from different substrates, insect-host, and localities from Colima, Mexico, were phylogenetically identified using the 5'end of translation elongation factor 1-α (5'TEF) and intergenic nuclear region MzFG543igs. Seven species were recognized, M. acridum (n = 26), M. pemphigi (n = 1), and within the PARB and MGT clades: M. anisopliae (N = 7; sensu stricto: n = 2; sensu lato: n = 5), M. brunneum (n = 2), M. guizhouense (n = 2), M. pingshaense (n = 2), and M. robertsii (n = 5). Twenty-nine SSR markers were developed for M. acridum; according to the analysis of 12 polymorphic SSR loci, M. acridum showed low genetic diversity, revealing five genotypes with a dominant one (n = 21). Based on the analysis of 13 specific SSR loci, 14 genotypes were identified within the PARB and MGT clades. This study contributes to generating valuable information about the community structure and genotypic diversity of Metharhizum species in the state of Colima, Mexico.

A new cryptogonimid (Digenea) from the Mayan cichlid, Cichlasoma urophthalmus (Osteichthyes: Cichlidae), in several localities of the Yucatán Peninsula, Mexico.

Oligogonotylus mayae n.sp. is described from the intestine of the Mayan cichlid Cichlasoma urophthalmus (Günther) in Ría Lagartos, Ría Celestún, and Estero Progreso, Yucatán State. This is the second species described for Oligogonotylus Watson, 1976, the other being O.manteri Watson, 1976. The new species is readily distinguished from O. manteri by the anterior extension of the vitelline follicles. In O. Manteri, Vitelline follicles are found entirely in the hindbody, extending posteriorly to mid-testicular level. Vitelline follicles in the new species extend from teh anterior margin of posterior testis to the region between the bentral sucker and the pharynx. comparison of approximately 1,850 bases of ribosomal DNA (ITS1, ITS2, 5.8S, and 28S), and 400 bases of cytochrome c oxidase subunit I (cox1) strongly supports the status of O. mayae as a new species, as compared to O. manteri collected from cichlids in other localities of Mexico, Belize, and Guatemala.

Description of a new species of Crassicutis Manter, 1936, parasite of Cichlasoma beani Jordan (Osteichthyes: Cichlidae) in Mexico, based on morphology and sequences of the ITS1 and 28S ribosomal RNA genes.

A new species of Crassicutis Manter, 1936 is described from the Sinaloan cichlid Cichlasoma beani (Jordan) (Osteichthyes: Cichlidae) in the upper Río Santiago basin. Crassicutis choudhuryi n. sp. differs from most of the other nominal species by having testes located in a symmetrical position. The only other species of the genus that includes some specimens exhibiting this trait is Crassicutis intermedius (Szidat 1954), a species found in 5 species of siluriforms and 1 species of characiform in South America. However, this species differs from Cr. choudhuryi n. sp. by having testes almost half of the size, and vitelline follicles extending anteriorly to the region between the acetabulum and the intestinal bifurcation. The new species is morphologically very similar to Crassicutis cichlasomae Manter, 1936, but clearly differs from this species because of the constantly symmetrical position of the testes. Additionally, Cr. choudhuryi n. sp. is found in the Santiago River basin on the Pacific slope of Mexico, parasitizing specifically the endemic Ci. beani that does not co-occur with any other cichlid. Cr. cichlasomae exhibits more hosts (about 25 species of cichlids only in Mexico) and a wider distribution range that extends from northeastern Mexico southward to Central America, Cuba, and Brazil. To corroborate that our specimens were not conspecific with Cr. cichlasomae, sequences of the internal transcribed spacer (ITS1) and the 28S ribosomal RNA genes of individuals from several populations (recently collected in southeastern Mexico) were obtained and compared to the species described herein. Sequence divergence (1.3% for the 28S and 4.0% for the ITS1) gives further support to the erection of a new species.

The phylogenetic position of Allocreadiidae (Trematoda: Digenea) from partial sequences of the 18S and 28S ribosomal RNA genes.

Species of Allocreadiidae are an important component of the parasite fauna of freshwater vertebrates, particularly fishes, and yet their systematic relationships with other trematodes have not been clarified. Partial sequences of the 18S and 28S ribosomal RNA genes from 3 representative species of Allocreadiidae, i.e., Crepidostomum cooperi, Bunodera mediovitellata, and Polylekithum ictaluri, and from 79 other taxa representing 78 families of trematodes obtained from GenBank, were used in a phylogenetic analysis to address the relationships of Allocreadiidae with other plagiorchiiforms/plagiorchiidans. Maximum parsimony and Bayesian analyses of combined 18S and 28S rRNA gene sequence data place 2 of the allocreadiids, Crepidostomum cooperi and Bunodera mediovitellata, in a clade with species of Callodistomidae and Gorgoderidae, which, in turn is sister to a clade containing Polylekithum ictaluri and representatives of Encyclometridae, Dicrocoelidae, and Orchipedidae, a grouping supported by high bootstrap values. These results suggest that Polylekithum ictaluri is not an allocreadiid, a conclusion that is supported by reported differences between its cercaria and that of other allocreadiids. Although details of the life cycle of callodistomids, the sister taxon to Allocreadiidae, remain unknown, the relationship of Allocreadiidae and Gorgoderidae is consistent with their larval development in bivalve, rather than gastropod, molluscs, and with their host relationships (predominantly freshwater vertebrates). The results also indicate that, whereas Allocreadiidae is not a basal taxon, it is not included within the suborder Plagiorchiata. No support was found for a direct relationship between allocreadiids and opecoelids either.