Game of clones: Is Wolbachia inducing speciation in a weevil with a mixed reproductive mode?

Parthenogenesis is widely distributed in Metazoa but it is especially frequent in weevils (Coleoptera, Curculionidae) with one fifth of all known cases. Previous studies have shown that in the tribe Naupactini parthenogenetic reproduction most likely originated with an infection of the endoparasitic bacterium Wolbachia pipientis. In particular, Pantomorus postfasciatus possess a mixed reproductive mode: some populations have males while in others they are absent, and females produce clones by thelytoky. To better understand this scenario, we studied the population structure and infection status in 64 individuals of P. postfasciatus from Argentina and Brazil. We sequenced two mitochondrial (COI and COII) and one nuclear (ITS-1) fragments and obtained two very divergent haplogroups, one corresponding to the sexual populations uninfected with Wolbachia, and another conforming a monophyletic parthenogenetic (or presumptively parthenogenetic) and infected clade. Each of these haplogroups was identified as an independently evolutionary unit by all species delimitation analyses accomplished: multilocus *BEAST and BP&P, and single locus GMYC and K/θ rule. Additionally, present evidence suggests that Wolbachia infection occurred at least twice in all-female populations of P. postfasciatus with two different bacterial strains. Speciation mediated by Wolbachia is a recently described phenomenon and the case of P. postfasciatus is the first known case in a diplo-diploid insect. A model that describes how thelytoky-inducing phenotypes of Wolbachia could generate new lineages is discussed.

Different changes in slope between the medial and lateral tibial plateau after open-wedge high tibial osteotomy.

PURPOSE: In contrast to radiographic measurements, MRI provides multiple slices of the knee joint in the sagittal plane, making it possible to assess the medial and lateral tibial slope separately. The purpose of this study is to investigate the effect of medial open-wedge high tibial osteotomy (MOWHTO) on bony and meniscal slope in the medial and lateral tibiofemoral compartments. It was hypothesised that greater changes on the medial tibial plateau would be observed compared with the lateral one. METHODS: A retrospective analysis of prospectively collected data was performed on pre- and post-operative MRIs from 21 patients (17 men and 4 women; age 52 ± 9 years). Inclusion criteria were varus alignment, medial compartment osteoarthritis and election for a primary MOWHTO. Each patient had a preoperative and a post-operative high-resolution MRI (3Tesla, Magnetom Trio, Siemens AG) at an average follow-up of 2.1 years. A previously published method was used to measure bony and meniscal slope for each compartment. The difference between pre- and post-operative tibial slope for both compartments was calculated and associated with the amount of frontal correction. RESULTS: There was a significant increase in bony tibial slope in both compartments following MOWHTO. When a change in bony tibial slope was detected in an individual patient, the change was larger in the medial compartment, with the average change also significantly greater (p < 0.01) in the medial compartment (2.4° ± 1.3°) compared with the lateral compartment (0.9° ± 1.1°). There was also a significant increase (p < 0.01) in the lateral tibial meniscal slope of 0.9° ± 1.4°, which was equivalent to the change in the bony lateral slope. The amount of frontal correction was not significantly associated with the amount of change in slope. CONCLUSIONS: The results suggest that the modification of the bony slope is larger in the medial compartment after MOWHTO, which is likely related to the location of the hinge on the lateral tibial cortex. These findings suggest that consideration of the medial and lateral tibial slope intra-operatively could be important to identify the optimal location of the hinge. However, further studies are required before recommending any modification to the surgical technique, as the potential clinical consequences of tibial slope alterations remain unknown. LEVEL OF EVIDENCE: IV.

Distribution of Taenia saginata metacestodes: a comparison of routine meat inspection and carcase dissection results in experimentally infected calves.

A comparison of techniques for detecting the presence of Cysticercus bovis in bovine carcasses was made by using carcass dissection and routine beef inspection guidelines. In the study, 28 calves were used after they were tested and found to be negative for the presence of anti-C. bovis serum antibodies and were inoculated orally with aliquots containing 6×10(4) Taenia saginata eggs. One hundred and twenty days after inoculation, the animals were slaughtered and a post mortem evaluation was done following Brazilian Federal Beef Inspection guidelines. This routine meat inspection was able to identify 71·42% of the assessed infected carcasses as being parasitized. This result implies that 28·58% of the infected carcasses would have been released as fit for human consumption since they would have been considered as free of C. bovis infection when using this method for carcass assessment. Only 3·07% of the total 2311 metacestodes present in the carcasses were identified by the conventional procedures of sanitary inspection. The assessment of different parts of the carcasses showed high infestation rates in shoulder clod (14·37%), head (11·21%), neck+chuck roll (8·05%), heart (7·75%) and top (inside) round (7·18%) which, together, were responsible for housing 48·51% of all the cysts found in the 24 beef cuts assessed. These numbers contrasted to the low incidence of cysts found in organs such as tongue (3·12%), diaphragm (1·69%) and esophagus (1·60%) which are usually described as predilection sites for the parasite.

Histopathology of the reproductive system of male sheep experimentally infected with Toxoplasma gondii.

The aim of this study was to investigate the histopathological changes in reproductive system (testicles, epididymis, seminal vesicles, and prostate) of small male ruminants after Toxoplasma gondii infection. Eight sheep were inoculated with T. gondii: group I, four sheep (2.0 × 10(5) P-strain oocysts); group II, four sheep (1.0 × 10(6) RH-strain tachyzoites); and group III, two uninfected sheep maintained as control. Infection with T. gondii was confirmed by seroconversion (indirect fluorescent antibody test-IgG) in all the infected animals beginning on post-inoculation day (PID) 7. On PID 70, all the animals were euthanized and tissue samples (testicles, epididymis, seminal vesicles, and prostate) were collected and processed for histological analysis. The main changes detected were a focal mononuclear interstitial inflammatory infiltrate in the prostate and seminal vesicles; diffuse testicular degeneration associated with calcification foci and a multifocal mononuclear interstitial inflammatory infiltrate; and a mononuclear interstitial infiltrate and focal necrotic areas of the muscle fibers surrounding the seminal vesicles. The histopathological findings of this work, along with the detection of T. gondii in the examined parenchyma tissues (immunohistochemistry) and the results obtained by other authors examining different tissues, suggest that histological changes diagnosed in the reproductive system of rams infected with T. gondii are strongly suggestive of toxoplasmatic infection.

Reliability analysis of visual examinations carried out by school-teachers and a dental assistant in the detection of dental caries.

AIM: This study aims to investigate the reliability of examinations performed by teachers and by a dental assistant in detection of cavitated surfaces. METHODS: A sample of 168 students, aged 5-14 years, attending a public school in Duque de Caxias, Rio de Janeiro, was examined by persons with three different training backgrounds: a dentist, a dental assistant, and schoolteachers. Examinations were performed in the school with the aid of a tongue blade under natural light. Kappa statistics were estimated to assess agreement between the observers. Sensitivity, specificity, and positive and negative predictive value tests helped validate observations performed by the group of laypersons. RESULTS: The findings suggest satisfactory agreement with the dentist, with kappa values of 0.730 and 0.781 for the teachers and the dental assistant, respectively. The absence of cavities was easily detected (specificity = 96%). More caution is required in positive results indicated by the teachers or the dental assistant because these were not always confirmed subsequently (sensitivity = 76%) by the dentist. CONCLUSION: The aid of untrained personnel in dental epidemiology was shown to be a valid alternative for a signposting role.

Prevalence of anti-Toxoplasma gondii antibodies in dairy cattle, dogs, and humans from the Jauru micro-region, Mato Grosso state, Brazil.

In order to determine the prevalence of anti-Toxoplasma gondii antibodies, 2000 serum samples from female dairy cattle belonging to 50 farms in the southwest of Mato Grosso state were analyzed by the indirect fluorescent antibody test (IFAT> or =64). Serum samples from 61 dogs (IFAT> or =40) and 116 humans (IFAT=40), all from the same farm, were also tested. Among these samples, 1420 (71.0%) cattle, 54 (88.5%) dogs, and 113 (97.4%) humans were seropositive for the infection. No significant differences (P> or =0.05) were observed for risk factors associated with the occurrence of toxoplasmosis in humans due to contaminated sources such as fresh milk, cheese/sausage, and contact with felines or other animals. The presence of felines can indicate the likelihood of a contaminated environment, posing a risk to the human population and other animals. The work presented herein is the first report to evaluate the seroprevalence of T. gondii in bovines from the Southwest region of the Mato Grosso state, Brazil.

Transplacental transmission of Toxoplasma gondii in reinfected pregnant female canines.

Twelve pregnant female canines, naturally infected with Toxoplasma gondii, were reinfected with T. gondii: three (GI) received tachyzoites subcutaneously (1.0 x 107), three (GII) were orally inoculated with oocysts (1.5 x 104), and six (GIII) were kept as a nonreinfected control group. All the reinfected female canines (GI and GII) miscarried or presented fetal death, while only one GIII female presented a stillborn in a litter of four pups (P < 0.01). Fever, lymphoadenopathy, miscarriage, and fetal death were the main clinical alterations observed. The highest serological titers detected through the indirect fluorescence antibody test (IFAT) were 1,024 (GI) and 4,096 (GII). In group III, the titers ranged between 64 and 256. By bioassays in mice, T. gondii was isolated in 17 organs of the reinfected adult canines, in 11 of the control group, and in 20 of the neonates. Positive immunostaining of cysts and/or tachyzoites were observed in 26 canine tissues (14 from GI and GII and ten from GIII). The agent was detected by immunohistochemistry in the encephalon of a neonate and in the spinal cord of a stillborn, thus, confirming that T. gondii infected canine fetuses, provoking miscarriages, even in bitches that presented primoinfection.

Endectocide activity of a new long-action formulation containing 2.25% ivermectin+1.25% abamectin in cattle.

The present work aimed to evaluate the endectocide activity of a new injectable long-action formulation, containing ivermectin (IVM) and abamectin (ABA). In each one of the four experiments performed, the following groups were formed: group I: 2.25% IVM (450 microg/kg)+1.25% ABA (250 microg/kg), group II: 3.15% IVM (630 microg/kg) and group III: control. Eighteen bovine naturally infected by gastrointestinal nematoda were selected for anthelmintic evaluation and necropsied on posttreatment day (PTD) 14 to estimate the total parasitic burden. For the Rhipicephalus (Boophilus) microplus field trial, 30 bovine were selected by means of counts of semi-engorged R. (B.) microplus and the therapeutic and residual efficacy evaluated by tick counts on PTDs 1, 3, 7, 14, 21, 28, 35, 42, 49, 56, 63, 70, 77, 84 and 91. In the stall test, 15 calves were artificially infested with 5000 R. (B.) microplus (Mozzo strain) larvae three times a week and daily collections of all the engorged female ticks detached from each calf were performed until the PTD 80. Forty bovine naturally infected with Dermatobia hominis larvae were selected and the number of larvae was counted by visual and tactile inspection on PTDs 3, 7, 14, 28, 35, 49, 63, 77, 91 and 105. In this trial, a formulation containing 1% doramectin (200 microg/kg) was also used. IVM+ABA formulation and 3.15% IVM eliminated four of the eight species of nematode identified. The anthelmintic efficacy of the avermectins association against Haemonchus placei, Cooperia spatulata and C. punctata was 89.64%, 98.84% and 97.69%, while 3.15% IVM achieved 30.98%, 84.79% and 75.56%, respectively. The two formulations evaluated showed reduced acaricide action on the PTD 1 and 3, reaching high efficacy percentages from PTD 14 onward. The IVM+ABA showed efficacy above 95% in the period between PTDs 21 and 49. In the stall test, it observed no difference (P>0.05) between the two formulations regarding the R. (B.) microplus counts during the entire evaluation period. IVM+ABA reduced the number of ticks from the PTD 1 to 77 (P<0.05) and 3.15% IVM reduced (P<0.05) the tick number from PTD 4 up to PTD 80. The three endectocides showed no difference (P>0.05) regarding the number of D. hominis larvae and prevented this parasite reestablishment until PTD 105. These results indicate that the IVM+ABA association showed higher anthelmintic activity and similar efficacy against arthropods to the formulation containing 3.15% IVM.

Absence of cardiovascular autonomic dysfunction and vagal pancreatic impairment in idiopathic achalasia of the oesophagus.

Extra-oesophageal autonomic dysfunction in idiopathic achalasia is not well documented, due to contradictory results reported. We aimed to study the cardiovascular and pancreatic autonomic function in patients with idiopathic achalasia. Thirty patients with idiopathic achalasia (16M/14F; 34.5 +/- 10.8 years) and 30 healthy volunteers (13M/17F; 34.8 +/- 10.7 years) were prospectively studied. Age >60 years and conditions affecting results of autonomic evaluation were excluded. Both groups underwent the sham feeding test and plasmatic levels of pancreatic polypeptide (PP) were determined by radioimmunoassay (basal, at 5, 10, 20 and 30 min). Cardiovascular parasympathetic (deep breathing, standing, Valsalva) and sympathetic function (postural decrease of systolic blood pressure, Handgrip test) were assessed. Statistical comparison of basal and increase levels of PP and parasympathetic/sympathetic cardiovascular parameters was performed between groups. Basal levels of PP were similar in controls and patients and maximum increase of PP during sham feeding test. A similar rate of abnormal cardiovascular tests was found between groups (P > 0.05). E/I ratio was the mostly impaired parameter (patients: 36.7% vs controls: 20%, P = 0.15, chi-squared test). Autonomic cardiovascular tests and pancreatic response to vagal stimulus are not impaired in patients with primary achalasia of the oesophagus.

Chemical basis for inter-colonial aggression in the stingless bee Scaptotrigona bipunctata (Hymenoptera: Apidae).

Inter-colonial aggression was tested using three colonies of Scaptotrigona bipunctata in a natural setting when their nests were moved and by artificial contact between individuals. Examination of the cuticular lipids of individuals from two colonies kept under identical conditions showed clear differences in their cuticular hydrocarbon profiles. The cuticular lipids were a mixture of hydrocarbons (saturated and unsaturated alkanes and alkenes) within the range of C23-C29. The use of multivariate analysis (PCA and discriminant analysis) showed that seven of the identified surface compounds are enough to separate workers from colonies A and B from each other.

Experimental toxoplasmosis in broiler chicks.

To evaluate chicken toxoplasmosis both as an economic and a public health subject, 84 broiler chicks of a commercial strain, 30 days old, were distributed into seven groups of 12 birds (three replications of four chicks) experimentally infected with three developing T. gondii stages of the P strain as follows: tachyzoites, intravenous (two groups: 5.0 x 10(5) and 5.0 x 10(6)), cysts, per os (two groups: 1.0 x 10(2) and 1.0 x 10(3)) and oocysts, per os (three groups: 5.0 x 10(2), 5.0 x 10(3) and 5.0 x 10(4)). Twelve chicks received only a placebo (control group). During the next 30 days the following parameters were estimated: productivity (weight gain and feed conversion), clinical signs, including rectal temperature and parasitemia (bioassay). No clinical signs suggesting toxoplasmosis were seen and no statistical differences on productivity standards were found in comparison between inoculated and control chicks. However, fowls inoculated with tachyzoites and oocysts occasionally showed hyperthermia. Some haematological changes were detected in fowls inoculated with T. gondii. Anatomo-histopathological changes were not observed. From 14 parasitemias detected, 35.7% appeared on the 5th day after inoculation and 57.1% of them resulted from oocysts inoculation. After 30-35 days all birds were slaughtered: fragments from 12 organs or tissues from each of them were subjected to artificial peptic digestion and after that injected into T. gondii antibody-free mice (IIFR). T. gondii was detected in brain (12), pancreas (five), spleen (five), retina (five), kidney (two), heart (four), proventriculus (three), liver (two), intestine (two), lung (one), and skeletal muscle (one). Similar to observations with parasitemia, from 42 T. gondii isolations, 59.5% came from chicks which had received oocysts. It can thus be inferred that the developing form, expelled by cats, is the most important for T. gondii chicken infection and that brain is the most infected organ in birds. Attention must be paid to the potential importance of chicken meat in public health, since T. gondii was isolated from skeletal and heart muscles.

Experimental toxoplasmosis in pregnant bitches.

Three Toxoplasma gondii free bitches (No. 3-5) were fed 15000 sporulated T. gondii oocysts at 56, 40 and 32 day of gestation and the outcome of the pregnancy was monitored. Two of the three dogs infected during pregnancy showed evidence of congenital infection and one aborted. Two control bitches not fed oocysts delivered eight uninfected healthy pups. This study demonstrated that T. gondii can be congenitally transmitted in dogs when bitches are infected during pregnancy.

Comparative efficacy evaluation of moxidectin gel and ivermectin paste against internal parasites of equines in Brazil.

A total of 24 male and female equines of mixed breed, 10-20 months of age and naturally infected with internal parasites was utilized in a controlled test to evaluate the efficacy of a moxidectin 2% gel formulation at the dosage of 0.4 mg moxidectin per kg of live weight and an ivermectin 1.87% commercial paste formulation at the dosage 0.2 mg ivermectin per kg applied orally. Animals were allocated into three groups of eight horses each based on pre-treatment eggs per gram (EPG) counts and treatments were randomized among the groups. One group was kept as untreated controls. One animal in the moxidectin-treated group died before the end of the trial from a cause unrelated to treatment leaving a total of seven animals in this group. Fecal egg counts were performed three times post-treatment and the number of parasites remaining in each animal was determined. Statistical analyses using geometric means were performed at the 1% level of significance. Both moxidectin and ivermectin preparations reduced initial EPG from a mean of 1600 to 0 on Days 5, 7 and at the end of the trial on Day 14. Efficacy percentages of moxidectin and ivermectin against immature and adult nematodes were as follows: Trichostrongylus axei, Parascaris equorum, Strongylus edentatus, S. vulgaris, Triodontophorus spp. and Gyalocephalus capitatus, 100% for both products; Habronema muscae 99.5 and 99.6%, respectively, Strongyloides westeri, 100 and 99.2%, respectively; Oxyuris equi, 99.6 and 100%, respectively; small strongyles, 99.7% for both products. Of the latter, the most numerous were: Cylicocyclus insigne, Cylicostephanus longibursatus and Cyathostomum catinatum. No Gasterophilus nasalis were found in horses from either treated group, while two of eight control horses had infections with thisparasite. Moxidectin showed greater efficacy (84.9%) than ivermectin (67.8%) against Strongylus vulgaris larvae found in the mesenteric artery aneurisms, but the difference was not statistically significant. Total parasite counts for both treated groups were significantly lower (p<0.01) than in the non-treated group. No significant differences were noted between moxidectin and ivermectin. Efficacy against the 30 nematode species found in this study was very evident for both products. As expected, neither moxidectin nor ivermectin was effective in controlling the tapeworm Anoplocephala perfoliata. No adverse reactions were observed during the experimental period.

Detection of IgG antibodies to Neospora caninum and Toxoplasma gondii in dogs examined in a veterinary hospital from Brazil.

A total of 163 dogs with neuromuscular, respiratory and/or gastrointestinal disorders, was admitted at the Veterinary Hospital, Federal University of Uberlândia, Brazil, and submitted to serology for Toxoplasma gondii and Neospora caninum. Assays for T. gondii included indirect haemagglutination (IHA), indirect fluorescent antibody (IFAT-Tg), immunoenzymatic (ELISA), and immunoblotting (IB-Tg). Assays for N. caninum included IFAT-Nc and immunoprecipitation (IP-Nc). Based on concordant results by three serological tests (IHA, IFAT-Tg and ELISA) for T. gondii, and divergent results further confirmed by IB-Tg for reactivity to TgSAG1, the 163 sera were divided into two groups: 59 (36%) Tg-seropositive samples and 104 (64%) Tg-seronegative samples. Antibodies to Neospora were detected in 11 (6.7%) out of 163 analyzed dog sera, with 5 (3.1%) samples reactive to both parasites (Tg+/Nc+), and 6 (3.7%) reactive only to Neospora (Tg-/Nc+). Antibodies only to T. gondii were found in 54 (33%) samples. Among the 11 Neospora-positive sera analyzed by IB-Tg, the five sera Tg+/Nc+ showed strong reactivity to Toxoplasma antigens, especially to TgSAG1 (p30). No reactivity was observed to TgSAG1 in the six samples Tg-/Nc+. By IP-Nc, two highly immunodominant antigens (29 and 35kDa proteins) were recognized by all 11 IFAT-Nc positive sera. Our results suggest that the infection by N. caninum can be concomitantly present in dogs from this area, although less common, and therefore should be considered in the differential clinical diagnosis with T. gondii in dogs presenting neuromuscular, respiratory and/or gastrointestinal disorders.

Experimental infection of bovines with oocysts of Toxoplasma gondii.

Calves aged 3 months were readily infected with oocysts and cysts of Toxoplasma gondii administered by oral route. Fever, respiratory distress, nasal discharge and hyperemia of the conjunctivas were the most significant clinical signs noted in the infected animals. Parasitemia was demonstrated in all infected calves. It occurred on different days and up to 62 days after the infection. Toxoplasma was demonstrated in tissues of all infected calves, and the organ most frequently parasitized was the lymph node. Parasitism of the retina was demonstrated in two calves. All infected animals had antibody against T. gondii in their serum. The Sabin-Feldman dye test and the indirect immunofluorescent test were both useful in detecting antitoxoplasma antibody.

To stay or not to stay: factors influencing family practice residents' choice of initial practice location.

BACKGROUND AND OBJECTIVES: This study examined what factors influence US third-year family practice residents' choice of location of their first practice. METHODS: A survey was developed by the members of Northeastern Ohio Network to assess the influence of factors related to family, education, geography, finances, the medical community, and others on the selection of practice location. Surveys were sent to all program directors listed in the American Academy of Family Physicians 1994 Directory of Family Practice Residency Programs to be distributed to all third-year family practice residents. The program directors indicated the type of residency program, the number of surveys distributed, and the number returned, then forwarded completed surveys to the investigators. RESULTS: Two hundred fifty-nine of the 380 programs (68%) with PGY-3 residents responded. Of the programs that responded, 1,012 residents (64%) of a possible 1,578 completed surveys. Estimated from the directory, 812 residents from non-responding programs did not complete the survey for an overall response rate among residents of 42% (1,012/[1,578+812]). Family-related items seem to have the most influence on choice of practice location and are more important to married people. More than 50% of residents want to practice in the same size community in which they grew up. Money-related factors were not rated as highly as expected. CONCLUSIONS: Several factors appear to be important to family practice residents when selecting their first practice location. Hospitals interested in retaining their graduates can maximize their success by concentrating on these areas.

Partial trisomy and partial monosomy of the distal long arm of chromosome 4: patient report and literature review.

We report the first patient with a partial trisomy and a partial monosomy of the long arm of chromosome 4: 46,XY, inv dup(4)(pter-->q32::q32-->q26), del(4)(q32-->qter). The boy died from a complex cardiac defect (monoventricle, monoatrium and truncus arteriosus) in combination with a diaphragmatic hernia. In addition he had preaxial polydactyly of the right hand. We compare the clinical features with data from the literature. The phenotype of the patient mainly resembles that in patients with a terminal deletion 4q32.

Elder abuse.

Elder abuse is a tragedy both for the individual and for society because it occupies a pivotal position in the family life cycle of violence. Great variety exists among victims, abusers, and situations; thus, no single model is adequate to explain cause and direct treatment. Primary care physicians must be alert to the possibility of elder abuse in their patients and aware of resources within their community for managing cases once identified. Federal laws and regulations must take a proactive, long-term approach to the solution of this problem and must respect the autonomy of competent elderly patients.

Serological findings during a measles outbreak occurring in a population with high vaccine coverage.

From March 1991 to April 1992, serum samples for IgM detection were collected from 112 clinical measles cases reported to the Health Department of Niterói, State of Rio de Janeiro. The positivity exceeded 90% for specimens collected from the 5th to the 29th day after the onset of the disease. After day 30 a decline in IgM detection was observed, although positivity has been detected up to 90 days after the onset of the symptoms. Forty-four patients (48.9%) with an IgM response had a history of prior measles vaccination. In 5 of the 22 measles-IgM negative cases the infection was due to other agents (rubella: 4 cases, dengue: 1 case). These results show that sensitivity of the test employed for confirming suspected measles cases is high, even in vaccinated patients.