Validation of the FAMACHA© system in South American camelids.

Haemonchus contortus resistant to multiple anthelmintics threaten the viability of the small ruminant industry in areas where this parasite is prevalent. In response to this situation, the FAMACHA© system was developed and validated for use with small ruminants as a way to detect clinical anemia associated with haemonchosis. Given that H. contortus and multiple anthelmintic resistance is a similar problem in camelids, the FAMACHA© system might also provide the same benefits. To address this need, a validation study of the FAMACHA© system was conducted on 21 alpaca and llama farms over a 2-year period. H. contortus was the predominant nematode parasite on 17 of the 21 farms (10 alpaca and 7 llama farms) enrolled in the study, based on fecal culture results. The FAMACHA© card was used to score the color of the lower palpebral (lower eye lid) conjunctiva on a 1-5 scale. Packed cell volume (PCV) values were measured and compared to FAMACHA© scores using FAMACHA© score cutoffs of ≥3 or ≥4 and with anemia defined as a PCV ≤15%, ≤17%, or≤20%. PCV was significantly associated with FAMACHA© score, fecal egg count (FEC), and body condition score (BCS), regardless of the FAMACHA© cutoff score or the PCV% chosen to define clinical anemia (p<0.01 in all cases). The use of FAMACHA© scores ≥3 and PCV ≥ 15% indicating anemia provided the best sensitivity (96.4% vs 92.9% for FAMACHA© ≥4), whereas FAMACHA scores ≥ 4 and PCV ≤20% provided the best specificity (94.2% vs 69.1% for FAMACHA© ≥3). The data from this study support the FAMACHA© system as a useful tool for detecting clinical anemia in camelids suffering from haemonchosis. Parameters for making treatment decisions based on FAMACHA© score in camelids should mirror those established for small ruminants.

Pathology of Haemonchus contortus in New World camelids in the southeastern United States: a retrospective review.

Most small ruminant farms in tropical climates are plagued by Haemonchus contortus, a hematophagous, abomasal parasite. Heavy burdens of this parasite can cause anemia, hypoproteinemia, weight loss, and mortality in susceptible animals. Haemonchus contortus is becoming a major health concern in New World camelids as well, namely llamas (Llama glama) and alpacas (Vicugna pacos), yet little research has been conducted regarding its prevalence or pathology in these species. Herein, we present a retrospective review of llamas and alpacas that were admitted to The University of Georgia Veterinary Teaching Hospital and Athens Diagnostic Laboratory between the years 2002 and 2013. Antemortem fecal egg count (FEC) estimates performed on 30 alpacas were negatively correlated with hematocrit, hemoglobin, and red blood cell count. Total protein was not significantly correlated with FEC. On postmortem examination, 55 of 198 camelids, including 2 from the aforementioned antemortem review, were infected with H. contortus, with llamas (42.6%) having a significantly higher infection rate than alpacas (22.2%). In 15.7% of the total cases, the parasite was the major cause of death. Common gross lesions included peritoneal, thoracic, and pericardial effusions, visceral pallor, subcutaneous edema, and serous atrophy of fat. Histologic lesions included centrilobular hepatic necrosis, hepatic atrophy, lymphoplasmacytic inflammation of the mucosa of the third gastric compartment (C3), extramedullary hematopoiesis in both the liver and spleen, and the presence of nematodes in C3. Our study emphasizes the importance of H. contortus diagnosis and herd monitoring in New World camelids, particularly llamas.

Auxin and strigolactone signaling are required for modulation of Arabidopsis shoot branching by nitrogen supply.

The degree of shoot branching is strongly affected by environmental conditions, such as nutrient availability. Here we demonstrate that nitrate limitation reduces shoot branching in Arabidopsis (Arabidopsis thaliana) both by delaying axillary bud activation and by attenuating the basipetal sequence of bud activation that is triggered following floral transition. Ammonium supply has similar effects, suggesting that they are caused by plant nitrogen (N) status, rather than direct nitrate signaling. We identify increased auxin export from active shoot apices, resulting in increased auxin in the polar auxin transport stream of the main stem, as a likely cause for the suppression of basal branches. Consistent with this idea, in the auxin response mutant axr1 and the strigolactone biosynthesis mutant more axillary growth1, increased retention of basal branches on low N is associated with a failure to increase auxin in the main stem. The complex interactions between the hormones that regulate branching make it difficult to rule out other mechanisms of N action, such as up-regulation of strigolactone synthesis. However, the proposed increase in auxin export from active buds can also explain how reduced shoot branching is achieved without compromising root growth, leading to the characteristic shift in relative biomass allocation to the root when N is limiting.

FHY3 promotes shoot branching and stress tolerance in Arabidopsis in an AXR1-dependent manner.

The transposase-related transcription factor FAR-RED ELONGATED HYPOCOTYL3 (FHY3) promotes seedling de-etiolation in far-red light, which is perceived by phytochrome A (phyA). In this role, FHY3 indirectly mediates the nuclear import of light-activated phyA, which triggers downstream transcriptional responses. Here, we present genetic evidence for additional roles of FHY3 in plant development and growth. New fhy3 alleles were isolated as suppressors of max2-1 (more axillary branching2-1), a strigolactone-insensitive mutant characterised by highly branched shoots. Branching suppression by fhy3, in both wild-type and max2-1 backgrounds, resulted from inhibition of axillary bud outgrowth. Additional roles in axillary meristem initiation were revealed in the revoluta (rev) fhy3 double mutant, with fhy3 enhancing rev mutant defects in axillary shoot meristem formation, as well as in floral meristem maintenance. fhy3 also affected embryonic and floral patterning with low penetrance, and displayed oxidative stress-related phenotypes of retarded leaf growth and of cell death. The fhy3 phenotypes of axillary bud outgrowth suppression and of stress-induced leaf growth retardation both required the AUXIN-RESISTANT1 gene, and are independent of phyA. Consistent with the recent discovery that FHY3 regulates many Arabidopsis promoters, our results suggest much wider roles for FHY3 in growth and development, either in concert with, or beyond, light signalling.

Comparison of three point-of-care blood glucose meters for use in adult and juvenile alpacas.

OBJECTIVE: To compare the performance of 3 point-of-care glucose meters in adult and juvenile alpacas with that of a laboratory-based analyzer. DESIGN: Evaluation study. ANIMALS: 35 adult alpacas and 21 juvenile alpacas. PROCEDURES: Whole blood samples obtained via jugular venipuncture were tested with all 3 point-of-care glucose meters; plasma samples were also tested with 1 of those meters. Glucose concentrations determined by use of the point-of-care meters were compared with results from the laboratory-based analyzer. RESULTS: Plasma glucose concentrations determined by use of the laboratory-based analyzer ranged from 36 to 693 mg/dL. Over the entire range of glucose concentrations tested, the Lin concordance correlation coefficient (agreement) was significant and excellent for all comparisons. Concordance decreased for 1 glucometer when testing whole blood samples over a narrower range of glucose concentrations (50 to 200 mg/dL). Bias was typically small (< 10 mg/dL) for 3 of the 4 comparisons but considerable for 1 meter with the use of whole blood. The limits of agreement were wide for all comparisons over the entire range of glucose concentrations tested but decreased to within acceptable limits when the narrower glucose range (50 to 200 mg/dL) was analyzed for 3 of the comparisons. For samples with a PCV < 25%, bias and the limits of agreement were greater for one of the meters tested. CONCLUSIONS AND CLINICAL RELEVANCE: Discrepancies between point-of-care glucose meters and reference techniques can be considerable in alpacas, emphasizing the importance of assessing individual meter performance in a target population.

One-stage prothrombin time, activated partial thromboplastin time, thrombin time, fibrin degradation product concentration, and antithrombin activity in healthy adult alpacas.

BACKGROUND: Alpacas are increasingly presented to veterinarians for evaluation and care. Reports of alpaca reference intervals for one-stage prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), concentration of fibrin degradation products (FDP), and antithrombin (AT) activities are scarce or nonexistent. OBJECTIVE: The aim of this study was to determine values for blood coagulation times (PT, aPTT, and TT), FDP concentrations, and AT activities in healthy adult alpacas. METHODS: Of blood samples collected from 35 clinically healthy adult alpacas via jugular venipuncture and placed into sodium citrate and FDP tubes, 29 samples were assayable for coagulation testing. PT, aPTT, and TT were determined by physical (mechanical) clot detection; AT activity was determined using a thrombin-specific chromogenic substrate end-point assay; and FDP concentrations were determined by the slide agglutination method. RESULTS: Median values and ranges (minimum-maximum) were determined for PT (8.7 seconds, 6.6-11.2 seconds), aPTT (17.3 seconds, 11.9-22.5 seconds), TT (10.2 seconds, 5.4-16.0 seconds), and AT activity (123.3%, 104.8-144.2%). The mean concentration of FDP was <8 µg/mL. CONCLUSION: These values for coagulation times, FDP concentration, and AT activity will provide a useful starting point in the diagnostic evaluation of ill adult alpacas.

Monitoring microemboli during cardiopulmonary bypass with the EDAC quantifier.

Gaseous emboli may be introduced into the bypass circuit both from the surgical field and during perfusionist interventions. While circuits provide good protection against massive air embolism, they do not remove gaseous microemboli (GME) from the bypass circuit. The purpose of this preliminary study is to assess the incidence of GME during bypass surgery and determine if increased GME counts were associated with specific events during bypass surgery. In 30 cases divided between 15 coronary artery bypass grafts and 15 valve repairs, GME were counted and sizedt the three locations on the bypass circuit using the EDAC" Quantifier (Luna Innovations, Roanoke, VA). A mean of 45,276 GME were detected after the arterial line filter during these 30 cases, with significantly more detected (p = .04) post filter during valve cases (mean = 72,137 +/- 22,113) than coronary artery bypass graft cases (mean = 18,416 +/- 7831). GME detected post filter were significantly correlated in time with counts detected in the venous line (p < .001). Specific events associated with high counts included the initiation of cardiopulmonary bypass, heart manipulations, insertion and removal of clamps, and the administration of drugs. Global factors associated with increased counts post filter included higher venous line counts and higher post reservoir/bubble trap counts. The mean number of microemboli detected during bypass surgery was much higher than reported in other studies of emboli incidence, most likely due to the increased sensitivity of the EDAC Quantifier compared to other detection modalities. The results furthermore suggest the need for further study of the clinical significance of these microemboli and what practices may be used to reduce GME incidence. Increased in vitro testing of the air handling capability of different circuit designs, along with more clinical studies assessing best clinical practices for reducing GME activity, is recommended.

Cell wall composition contributes to the control of transpiration efficiency in Arabidopsis thaliana.

To identify loci in Arabidopsis involved in the control of transpirational water loss and transpiration efficiency (TE) we carried out an infrared thermal imaging-based screen. We report the identification of a new allele of the Arabidopsis CesA7 cellulose synthase locus designated AtCesA7(irx3-5) involved in the control of TE. Leaves of the AtCesA7(irx3-5) mutant are warmer than the wild type (WT). This is due to reduced stomatal pore widths brought about by guard cells that are significantly smaller than the WT. The xylem of the AtCesA7(irx3-5) mutant is also partially collapsed, and we suggest that the small guard cells in the mutant result from decreased water supply to the developing leaf. We used carbon isotope discrimination to show that TE is increased in AtCesA7(irx3-5) when compared with the WT. Our work identifies a new class of genes that affects TE and raises the possibility that other genes involved in cell wall biosynthesis will have an impact on water use efficiency.

Strigolactones enhance competition between shoot branches by dampening auxin transport.

Strigolactones (SLs), or their derivatives, were recently demonstrated to act as endogenous shoot branching inhibitors, but their biosynthesis and mechanism of action are poorly understood. Here we show that the branching phenotype of mutants in the Arabidopsis P450 family member, MAX1, can be fully rescued by strigolactone addition, suggesting that MAX1 acts in SL synthesis. We demonstrate that SLs modulate polar auxin transport to control branching and that both the synthetic SL GR24 and endogenous SL synthesis significantly reduce the basipetal transport of a second branch-regulating hormone, auxin. Importantly, GR24 inhibits branching only in the presence of auxin in the main stem, and enhances competition between two branches on a common stem. Together, these results support two current hypotheses: that auxin moving down the main stem inhibits branch activity by preventing the establishment of auxin transport out of axillary branches; and that SLs act by dampening auxin transport, thus enhancing competition between branches.

Efficacy of anthelmintics on South American camelid (llama and alpaca) farms in Georgia.

The number of South American camelid (SAC; llama and alpaca) farms is growing in the southeastern United States, and infection with gastrointestinal nematodes (GIN) is a major health concern in this region. There is widespread resistance to anthelmintic remedies in small ruminants (sheep and goats), but a paucity of information on llamas and alpacas. Anthelmintic resistance was evaluated on three SAC farms (two llama; one alpaca) in Georgia in the southern United States using fecal egg count reduction (FECR) tests. For each farm, animals were randomly assigned to 1 of 5 treatment groups based on initial fecal egg count (FEC) and number of animals available (2-5 groups, n=9-11 per treatment). Ivermectin (IVM, subcutaneous injection; 0.3mg/kg body weight (BW)) and a control group were tested on an alpaca farm, and fenbendazole (FBZ, oral; 10mg/kg BW; two farms), moxidectin (MOX oral; 0.2mg/kg BW; two farms), and levamisole (LEV, oral; 8 mg/kg BW; one farm) were added for the llama farms. Anthelmintic efficacy was determined by comparing FEC of treatment and control animals 14 days post-treatment, with resistance evaluated using the World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines. Based upon these guidelines, there was GIN resistance to IVM in both llamas and alpacas in Georgia and to FBZ on both llama farms where this drug was tested. There was MOX resistance on one llama farm using the FECR test, while there was no resistance to LEV detected in this study. These data demonstrate a serious emerging problem in the United States of llama and alpaca GIN resistant to drugs from two of the three major anthelmintic classes.

Young women's continued use of oral contraceptives over other hormonal methods: findings from a qualitative study.

BACKGROUND: Long-acting reversible contraceptives (LARC) have become more commonly promoted in the UK, but most young women still rely on the contraceptive pill. Here, we describe young women's accounts of hormonal contraceptive use to explore why this might be the case. METHODS: In-depth interviews with twenty 20-year-old women from eastern Scotland in the UK. RESULTS: All but one woman reported use of the pill. It was the method they expected to use, sought out, and received. Belief in the pill's efficacy was maintained even when knowledge or experience of failure suggested otherwise. Only four women reported using alternative hormonal methods and only did so after experiencing unmanageable problems with the pill (side effects or forgetting to take it). All then discontinued use because of weight gain or dislike of menstrual suppression. CONCLUSIONS: Attempts to promote LARC must address these issues. Pill use can be unproblematic if managed well, and should continue to be promoted as an appropriate contraceptive for young women.

Young women and limits to the normalisation of condom use: a qualitative study.

Encouraging condom use among young women is a major focus of HIV/STI prevention efforts but the degree to which they see themselves as being at risk limits their use of the method. In this paper, we examine the extent to which condom use has become normalised among young women. In-depth interviews were conducted with 20 year old women from eastern Scotland (N=20). Purposive sampling was used to select a heterogeneous group with different levels of sexual experience and from different social backgrounds. All of the interviewees had used (male) condoms but only three reported consistent use. The rest had changed to other methods, most often the pill, though they typically went back to using condoms occasionally. Condoms were talked about as the most readily available contraceptive method, and were most often the first contraceptive method used. The young women had ingrained expectations of use, but for most, these norms centred only on their new or casual partners, with whom not using condoms was thought to be irresponsible. Many reported negative experiences with condoms, and condom dislike and failure were common, lessening trust in the method. Although the sexually transmitted infection (STI) prevention provided by condoms was important, this was seen as additional, and secondary, to pregnancy prevention. As the perceived risks of STIs lessened in relationships with boyfriends, so did condom use. The promotion of condoms for STI prevention alone fails to consider the wider influences of partners and young women's negative experiences of the method. Focusing on the development of condom negotiation skills alone will not address these issues. Interventions to counter dislike, method failure, and the limits of the normalisation of condom use should be included in STI prevention efforts.

Young women's perceptions of pregnancy risk and use of emergency contraception: findings from a qualitative study.

BACKGROUND: Advance provision of emergency contraception (EC) has increased use but not impacted on pregnancy or abortion rates. Here we describe young women's EC use and experiences of unprotected sex to explore why this difference occurs. METHODS: In-depth interviews with twenty 20-year-old women from eastern Scotland. RESULTS: The majority (16) had used EC; 10 reported some experience of unprotected sex. EC use followed contraceptive failure and unexpected or unplanned, but not frequent, unprotected sex. Acknowledging the need for EC requires recognition of pregnancy risk. Those reporting frequent unprotected sex misperceived their pregnancy risk and did not use EC. This group was from socially disadvantaged backgrounds, and all became pregnant. CONCLUSIONS: EC remains an important "backup" contraceptive and should continue to be widely available. With high levels of unprotected sex, nonuse of EC and unintended pregnancies, further efforts are required to improve the sexual and reproductive health outcomes of disadvantaged young women.

Limits to modern contraceptive use among young women in developing countries: a systematic review of qualitative research.

BACKGROUND: Improving the reproductive health of young women in developing countries requires access to safe and effective methods of fertility control, but most rely on traditional rather than modern contraceptives such as condoms or oral/injectable hormonal methods. We conducted a systematic review of qualitative research to examine the limits to modern contraceptive use identified by young women in developing countries. Focusing on qualitative research allows the assessment of complex processes often missed in quantitative analyses. METHODS: Literature searches of 23 databases, including Medline, Embase and POPLINE(R), were conducted. Literature from 1970-2006 concerning the 11-24 years age group was included. Studies were critically appraised and meta-ethnography was used to synthesise the data. RESULTS: Of the 12 studies which met the inclusion criteria, seven met the quality criteria and are included in the synthesis (six from sub-Saharan Africa; one from South-East Asia). Sample sizes ranged from 16 to 149 young women (age range 13-19 years). Four of the studies were urban based, one was rural, one semi-rural, and one mixed (predominantly rural). Use of hormonal methods was limited by lack of knowledge, obstacles to access and concern over side effects, especially fear of infertility. Although often more accessible, and sometimes more attractive than hormonal methods, condom use was limited by association with disease and promiscuity, together with greater male control. As a result young women often relied on traditional methods or abortion. Although the review was limited to five countries and conditions are not homogenous for all young women in all developing countries, the overarching themes were common across different settings and contexts, supporting the potential transferability of interventions to improve reproductive health. CONCLUSION: Increasing modern contraceptive method use requires community-wide, multifaceted interventions and the combined provision of information, life skills, support and access to youth-friendly services. Interventions should aim to counter negative perceptions of modern contraceptive methods and the dual role of condoms for contraception and STI prevention should be exploited, despite the challenges involved.

Prevalence of anthelmintic resistance on sheep and goat farms in the southeastern United States.

OBJECTIVE: To determine prevalence of anthelmintic resistance on sheep and goat farms in the southeastern United States. DESIGN: Cross-sectional study. ANIMALS: Sheep and goats from 46 farms in 8 southern states, Puerto Rico, and St Croix in the US Virgin Islands. PROCEDURES: Parasite eggs were isolated from fecal samples, and susceptibility to benzimidazole, imidathiazole, and avermectin-milbemycin anthelmintics was evaluated with a commercial larval development assay. RESULTS: Haemonchus contortus was the most common parasite on 44 of 46 farms; Trichostrongylus colubriformis was the second most commonly identified parasite. Haemonchus contortus from 45 (98%), 25 (54%), 35 (76%), and 11 (24%) farms were resistant to benzimidazole, levamisole, ivermectin, and moxidectin, respectively. Resistance to all 3 classes of anthelmintics was detected on 22 (48%) farms, and resistance to all 3 classes plus moxidectin was detected on 8 farms (17%). CONCLUSIONS AND CLINICAL RELEVANCE: Findings provided strong evidence that anthelmintic resistance is a serious problem on small ruminant farms throughout the southeastern United States. Owing to the frequent movement of animals among regions, the prevalence of resistance in other regions of the United States is likely to also be high. Consequently, testing of parasite eggs for anthelmintic resistance should be a routine part of parasite management on small ruminant farms.

Sexual risk behaviour and knowledge of HIV status among community samples of gay men in the UK.

OBJECTIVES: Undiagnosed infection presents a potential risk for the onward transmission of HIV and denies men early interventions for their health. Little is known about the differences between men who are, and are not, aware of their HIV status in the UK. METHODS: Cross-sectional surveys of men in commercial gay venues in London, Brighton, Manchester, Glasgow, and Edinburgh (2003-2005). Anonymous, self-completion questionnaires, and oral fluid samples (tested for HIV antibodies) were obtained from 3672 men (61% response rate). RESULTS: Of 3501 men with a confirmed positive or negative oral fluid result, 318 were HIV positive (9.1%). Of these, 131 (41.2%) were undiagnosed; 81.1% of men with undiagnosed HIV had previously tested (92.2% tested negative; the remainder did not know the result); 62.3% still thought that they were negative. Undiagnosed and diagnosed men reported greater sexual risk and sexually transmitted infections than HIV-negative men. Compared with HIV-negative men, the adjusted odds ratio of unprotected anal intercourse with two or more partners was higher among undiagnosed men (odds ratio 2.21, 95% confidence interval 1.17-4.20), but highest among diagnosed men (odds ratio 6.80, 95% confidence interval 4.39-10.52). CONCLUSION: A high proportion of the HIV-positive men were undiagnosed and not receiving benefits of clinical care, but sexual risk and sexually transmitted infections were highest among men who were aware of their HIV-positive status. Clinics should proactively offer testing to reduce undiagnosed HIV, target repeat testing at high-risk men who have previously tested negative, and initiate evidence-based behavioural interventions to reduce sexual risk among men living with diagnosed HIV as well as those testing negative.

What explains between-school differences in rates of sexual experience?

BACKGROUND: Schools have the potential to influence their pupils' behaviour through the school's social organisation and culture, as well as through the formal curriculum. This paper provides the first attempt to explain the differences between schools in rates of reported heterosexual sexual experience amongst 15 and 16 year olds. It first examined whether variations in rates of sexual experience remained after controlling for the known predictors of sexual activity. It then examined whether these residuals, or 'school effects', were attributable to processes within the school, or were more likely to reflect characteristics of the neighbourhood. METHODS: Longitudinal survey data from 4,926 pupils in 24 Scottish schools were linked to qualitative and quantitative data on school processes including quality of relationships (staff-pupil, etc), classroom discipline, organisation of Personal and Social Education, school appearance and pupil morale. Multi-level modelling was used to test a range of models and the resulting 'school effects' were then interpreted using the process data. RESULTS: Overall, 42% of girls and 33% of boys reported experience of sexual intercourse, with rates by school ranging from 23% to 61%. When individual socio-economic and socio-cultural factors were taken into account the school variation dropped sharply, though pupils' attitudes and aspirations had little effect. There was very little correlation between boys' and girls' rates of sexual experience by school, after controlling for known predictors of sexual activity. Girls were more influenced by individual socio-economic factors than boys. School-level socio-economic factors were predictive even after taking account of individual socio-cultural factors, suggesting that the wider socio-economic environment further influenced young people's sexual experience. CONCLUSION: Importantly, school processes did not explain the variation between schools in sexual experience. Rather, the variation may have been due to neighbourhood culture.

Interactions between axillary branches of Arabidopsis.

Studies of apical dominance have benefited greatly from two-branch assays in pea and bean, in which the shoot system is trimmed back to leave only two active cotyledonary axillary branches. In these two-branch shoots, a large body of evidence shows that one actively growing branch is able to inhibit the growth of the other, prompting studies on the nature of the inhibitory signals, which are still poorly understood. Here, we describe the establishment of two-branch assays in Arabidopsis, using consecutive branches on the bolting stem. As with the classical studies in pea and bean, these consecutive branches are able to inhibit one another's growth. Not only can the upper branch inhibit the lower branch, but also the lower branch can inhibit the upper branch, illustrating the bi-directional action of the inhibitory signals. Using mutants, we show that the inhibition is partially dependent on the MAX pathway and that while the inhibition is clearly transmitted across the stem from the active to the inhibited branch, the vascular connectivity of the two branches is weak, and the MAX pathway is capable of acting unilaterally in the stem.