Longitudinal Associations Between Trauma Exposure and Executive Functions in Children: Findings from a Dutch Birth Cohort Study.

This study is the first to distinguish two possible predictive directions between trauma exposure and executive functioning in children in a community sample. The sample consists of 1006 children from two time points with a seven years' time interval of a longitudinal Dutch birth cohort study, the ABCD-study (Van Eijsden et al., 2011). We analyzed the longitudinal associations between trauma exposure and executive functioning using structural equation modeling. The results demonstrated that (after controlling for prenatal substance exposure and mothers' educational level) trauma exposure before age 5 is predictive of poorer executive functioning at age 12 and trauma exposure between age 6 and 12. However, the association between executive functioning at age 5 and trauma exposure between age 6 and 12 was not statistically significant. Our results indicate that early life trauma exposure has a long term impact on later executive functioning and not the other way around. On top of that, trauma exposure seems to accumulate across childhood when children are exposed to a traumatic event before the age of 5. When looking at the potential moderating role of parenting behavior we found no evidence for such a moderating effect of parenting behavior. Our findings showed that children exposed to trauma early in life may experience problems in executive functioning later in life and they seem at higher risk for cumulative trauma exposure. Clinical practice should take this into account in both the way they provide (early) mental health care and in prevention and recognition of early trauma exposure.

An atypical presentation of a mesenteric Meckel's diverticulum in a 7-year-old warmblood mare: case report.

BACKGROUND: Meckel's diverticula are a rare cause of small intestinal strangulation, diagnosed at laparotomy or necropsy. This congenital anomaly of the gastrointestinal tract originates from a remnant of the vitelline duct. In reported equine cases, they present as a full-thickness diverticulum on the antimesenteric border of the distal jejunum or proximal ileum. CASE PRESENTATION: On laparotomy a Meckel's diverticulum positioned at the mesenteric side was found to be the cause of small intestinal strangulation. This position is very uncommon and to the best knowledge of the authors there is no unambiguous description of another case. CONCLUSIONS: Meckel's diverticula should be on the list of differential diagnoses in cases of small intestinal strangulation. As in humans, equine Meckel's diverticula can have the standard antimesenteric as well as a more exceptional mesenteric location. This case adds to the series of anecdotal reports of anomalies with regard to Meckel's diverticula in the horse.

Interaction and imbalance between indispensable amino acids in young piglets.

Lowering protein level in diets for piglets urge to have knowledge on the piglet's requirements for essential amino acids (AA) and their interactions. The present studies aimed to determine the interaction between the dietary level of valine (Val) and tryptophan (Trp) and the effect of AA imbalance at two levels of dietary Val on the growth performance of post-weaning piglets. In Experiment 1 (duration 4 weeks), the effects of supplementation of free l-Val (1.0 g/kg) and/or l-Trp (0.5 g/kg) in a low-CP diet (CP 17.7%), marginal in Trp and Val, was studied in a 2×2 factorial design and using an additional reference treatment (CP 19.5%). In Experiment 2 (duration 5 weeks), the influence of a stepwise increase in excess supply of isoleucine (Ile), histidine (His) and leucine (Leu), up to 10, 10% and 30% relative to their requirement values respectively, was evaluated at 60% or 70% standardized ileal digestible (SID) Val relative to SID lysine, using a 3×2 factorial design. In Experiment 1, over the whole experimental period, feed intake (FI) was affected by dietary Trp level (P<0.05) and feed conversion ratio (FCR) by both the level of Trp and Val in the diet (both P<0.05). Increasing Trp level increased FI and decreased FCR while increasing dietary Val level reduced FI and increased FCR. For BW gain (BWG), there was an interaction between dietary level of Trp and Val (P<0.05). Valine supplementation decreased BWG using a diet marginal in Trp, whereas it increased BWG when using a Trp sufficient diet. Piglets fed the low-CP diet with adequate levels of Val and Trp showed at least same performance compared to piglets fed the high CP reference diet. In Experiment 2, increasing dietary Val improved FI and BWG (P<0.001) and tended to improve FCR. Dietary AA excess for Ile, His and Leu reduced FI and BWG (P<0.05) and only affected FCR (P<0.01) in the 1st week of the study. Dietary level of Val and AA excess did not show interactive effects, except for FCR over the final 2 weeks of the study (P<0.05). In conclusion, an interaction exists between dietary supply of Val and Trp on the zootechnical performance of post-weaning piglets and dietary AA excess for Ile, Leu and His, reduces growth performance of piglets in low-protein diets, independent of the dietary level of Val.

Expression of p53, Ki67, EcPV2- and EcPV3 DNA, and viral genes in relation to metastasis and outcome in equine penile and preputial squamous cell carcinoma.

REASONS FOR PERFORMING STUDY: Equine penile and preputial squamous cell carcinoma (SCC) is a potentially lethal disease of which little is known regarding the relationship between tumour characteristics and prognosis. OBJECTIVES: To assess the relationship between tumour differentiation grade (tumour subtype), presence of papillomaviruses, expression of viral genes (E2, E6, L1), nuclear proteins p53 and Ki67 and metastasis in equine penile and preputial SCC and to assess the relationship of tumour subtype, presence of papillomavirus type 2, p53 and Ki67 with survival. STUDY DESIGN: Retrospective case-control study using archived material. METHODS: Samples (n = 103) from 87 horses with penile and/or preputial intraepithelial neoplasia (PIN), papilloma or SCC and corresponding case files were evaluated. Tumours were graded microscopically and p53 and Ki67 expression evaluated immunohistochemically. Equine papillomavirus (EcPV) types 2 and 3 DNA was detected by conventional PCR. Real-time PCR was used for quantification of E2, E6 and L1 mRNA. RESULTS: Equine papillomavirus type 2 DNA was detected in 89.4% and EcPV3 in 1.5% of horses. No differences in quantitative expression of E2, E6 and L1 oncogenes between subtypes were found. Expression of p53 and occurrence of metastasis were positively correlated to a less differentiated subtype (r = 0.429, P<0.001 and r = 0.769, P = 0.001, respectively). Differences in survival between subtypes were significant (log Rank P<0.001); horses with less differentiated tumours were more likely to die of the disease (papilloma 8.3%; G1 26.1%; G2 26.3%; G3 63.3%). CONCLUSIONS: In equine penile and preputial SCC, tumour grading is an important prognosticator for survival and a predictor for presence of metastases. Expression of p53 and Ki67 and presence or expression of EcPV2 and EcPV3 do not appear to be important prognosticators.

Expression of cyclo-oxygenases-1 and -2, and microsomal prostaglandin E synthase-1 in penile and preputial papillomas and squamous cell carcinomas in the horse.

REASONS FOR PERFORMING STUDY: Penile and preputial papilloma and squamous cell carcinoma (SCC) are commonly diagnosed in horses. Papillomas have the potential to progress to potentially lethal SCC. Knowledge of pathogenetic mechanisms may help in prevention and definition of treatment targets. STUDY DESIGN: Retrospective study using archived material. OBJECTIVES: To determine the expression of cyclo-oxygenase 1 (COX-1), cyclo-oxygenase 2 (COX-2) and microsomal prostaglandin E synthase-1 (mPGES-1) in penile and preputial normal tissue, papilloma and SCC in horses, and whether expression of these enzymes is influenced by degree of inflammation and differentiation grade. METHODS: Tumour differentiation grade, degree of inflammation and COX-1, COX-2 and mPGES-1 expression in 75 formalin-fixed paraffin embedded samples of penile and preputial papilloma and SCC of 68 horses were investigated by histopathology and immunohistochemistry. RESULTS: Inflammation was more prominent in SCC compared with papilloma. No correlation between expression of COX-1 or COX-2 and inflammation was found. Expression of mPGES-1 was weakly correlated with inflammation. Expression of COX-1, COX-2 and mPGES-1 was found in 42.6%, 50.7% and 96.0% of lesions respectively, but less than 1% of cells were immunopositive for COX-1 and COX-2 in 59.4% and 84.2% of cases respectively. Expression of COX-1 was moderately negatively correlated with differentiation grade, COX-2 was not correlated and mPGES-1 was poorly negatively correlated. CONCLUSIONS: Expression of COX-1 and COX-2 in penile and preputial SCC in the horse is poor and COX inhibitors may thus be of little value for prevention or treatment. Microsomal PGES-1 is more prominently expressed in well-differentiated tissue compared with poorly differentiated tissue. Further research on the role of mPGES-1 in carcinogenesis is needed to assess its potential use as a treatment target. Knowledge of arachidonic pathway enzyme expression and their role in equine penile and preputial carcinogenesis may help in developing preventive and therapeutic strategies.

Treatment and long-term follow-up of distichiasis, with special reference to the Friesian horse: a case series.

REASONS FOR PERFORMING STUDY: Distichiasis is very uncommon in horses and treatment options and outcome remain unknown. OBJECTIVES: To describe treatment and long-term follow-up of distichiasis in the horse. STUDY DESIGN: Retrospective clinical study. METHODS: Case records of horses admitted for diagnosis and treatment of uni- or bilateral distichiasis between 2003 and 2012 were reviewed. Long-term (>6 months after last treatment) follow-up was by telephone conversation with the owner or referring veterinarian. Data were analysed using a Chi-squared test. RESULTS: Eighteen cases were identified, of which 17 were Friesians. All horses had signs of corneal irritation or ulceration concurrent with distichiasis. In total, 29 eyes were affected with distichiasis of which 23 had clinical signs. All corneas were stained with fluorescein and 13 had fluorescein positive defects (45%). The lower eyelid was affected more often. Two different electrocautery devices were used for treatment. The complications were slight scarring of the eyelids (100%) and clinical recurrence (54%). Successful resolution of the clinical signs after one treatment occurred in 46% of the horses and in 77% of the horses satisfactory long-term results were found after a mean of 2 treatments (range: 1- 6 treatments). There was significant correlation between number of hairs and recurrence (P<0.002). CONCLUSIONS: There is a breed predisposition of distichiasis in the Friesian horse. In Friesian horses that present with uni- or bilateral corneal irritation or corneal ulceration, careful examination of the eyelid margins should be performed to diagnose distichiasis. Electrocautery is a valid and successful therapeutic modality which is minimally invasive, inexpensive, has a low complication rate and is easy to perform in the standing sedated horse. The recurrence rate is low in horses with less than 5 distichiae.

Escherichia coli contamination and health aspects of soil and tomatoes (Solanum lycopersicum L.) subsurface drip irrigated with on-site treated domestic wastewater.

Faecal contamination of soil and tomatoes irrigated by sprinkler as well as surface and subsurface drip irrigation with treated domestic wastewater were compared in 2007 and 2008 at experimental sites in Crete and Italy. Wastewater was treated by Membrane Bio Reactor (MBR) technology, gravel filtration or UV-treatment before used for irrigation. Irrigation water, soil and tomato samples were collected during two cropping seasons and enumerated for the faecal indicator bacterium Escherichia coli and helminth eggs. The study found elevated levels of E. coli in irrigation water (mean: Italy 1753 cell forming unit (cfu) per 100 ml and Crete 488 cfu per 100 ml) and low concentrations of E. coli in soil (mean: Italy 95 cfu g(-1) and Crete 33 cfu g(-1)). Only two out of 84 tomato samples in Crete contained E. coli (mean: 2700 cfu g(-1)) while tomatoes from Italy were free of E. coli. No helminth eggs were found in the irrigation water or on the tomatoes from Crete. Two tomato samples out of 36 from Italy were contaminated by helminth eggs (mean: 0.18 eggs g(-1)) and had been irrigated with treated wastewater and tap water, respectively. Pulsed Field Gel Electrophoresis DNA fingerprints of E. coli collected during 2008 showed no identical pattern between water and soil isolates which indicates contribution from other environmental sources with E. coli, e.g. wildlife. A quantitative microbial risk assessment (QMRA) model with Monte Carlo simulations adopted by the World Health Organization (WHO) found the use of tap water and treated wastewater to be associated with risks that exceed permissible limits as proposed by the WHO (1.0 × 10(-3) disease risk per person per year) for the accidental ingestion of irrigated soil by farmers (Crete: 0.67 pppy and Italy: 1.0 pppy). The QMRA found that the consumption of tomatoes in Italy was deemed to be safe while permissible limits were exceeded in Crete (1.0 pppy). Overall the quality of tomatoes was safe for human consumption since the disease risk found on Crete was based on only two contaminated tomato samples. It is a fundamental limitation of the WHO QMRA model that it is not based on actual pathogen numbers, but rather on numbers of E. coli converted to estimated pathogen numbers, since it is widely accepted that there is poor correlation between E. coli and viral and parasite pathogens. Our findings also stress the importance of the external environment, typically wildlife, as sources of faecal contamination.

Penile and preputial tumours in the horse: literature review and proposal of a standardised approach.

Penile and preputial tumours are not uncommon in the horse, but can cause discomfort and lead to serious complications. Several types of tumour of the male external genitalia have been described. The most common type is the squamous cell carcinoma (SCC), which is found mainly in older horses. Reports of a breed predilection for penile tumour formation are equivocal, but castration, coat colour, poor hygiene and various infectious agents have all been suggested to predispose to the development of some types of tumour (e.g. SCC, papilloma and melanoma). Careful assessment of the primary tumour is an important first step in the design of an optimal treatment protocol. Invasiveness, differentiation grade, tumour size and presence of metastases are all relevant to the decision to pursue additional diagnostic procedures or specific treatment options. To date, no standard protocol has been reported for the approach to penile tumours in the horse and treatments range from minimally invasive therapies (e.g. topical use of 5-fluorouracil) to radical surgical interventions (e.g. en bloc penile and preputial resection with penile retroversion). Completeness of removal of the neoplasm and therefore risk of recurrence is highly dependent on the type of therapy chosen. However, the size and histopathological features of the primary tumour are also important factors with respect to the likelihood of recurrence. This review describes the most common penile and preputial neoplasms in the horse, and outlines a standard protocol aimed at arriving at a specific diagnosis and tailoring the therapeutic approach accordingly.

Effect of feeding a reduced-starch diet with or without amylase addition on lactation performance in dairy cows.

The objective of this study was to determine lactation performance responses of high-producing dairy cows to a reduced-starch diet compared with a normal-starch diet and to the addition of exogenous amylase to the reduced-starch diet. Thirty-six multiparous Holstein cows (51+/-22 DIM and 643+/-49kg of body weight at trial initiation) were randomly assigned to 1 of 3 treatments in a completely randomized design: a 3-wk covariate adjustment period during which the cows were fed the normal-starch diet, followed by a 12-wk treatment period during which the cows were fed their assigned treatment diets. The normal-starch TMR did not contain exogenous amylase (NS-). The reduced-starch diets, formulated by partially replacing corn grain with soy hulls, were fed without (RS-) and with (RS+) exogenous amylase added to the TMR. Starch and NDF concentrations averaged 27.1 and 30.6%, 21.8 and 36.6%, and 20.7 and 36.6% (dry matter basis) for the NS-, RS-, and RS+ diets, respectively. Dry matter intake for cows fed the RS- diet was 2.4 and 3.2kg/d greater than for cows fed the NS- and RS+ diets, respectively. Intake of NDF ranged from 1.19 to 1.52% of body weight among the treatments, with the RS- diet being 28% greater than the NS- diet and 13% greater than the RS+ diet. Milk yield averaged 50.4kg/d and was unaffected by treatment. Fat-corrected milk yield was 2.9kg/d greater for cows fed the RS- diet than for cows fed the NS- diet. Body weight and body condition score measurements were unaffected by treatment. Fat-, solids-, and energy-corrected milk feed conversions (kilograms/kilogram of DMI) were 12 to 13% greater for cows fed the RS+ diet than for cows fed the RS- diet. Dry matter and nutrient digestibilities were lowest for cows fed the NS- diet and greatest for cows fed the RS+ diet, and were greater for cows fed the RS+ diet than for cows fed the RS- diet, with the exception of starch digestibility, which was similar. Greater conversion of feed to milk for dairy cows fed reduced-starch diets that include exogenous amylase may offer potential for improving economic performance.

Penile and preputial tumours in the horse: a retrospective study of 114 affected horses.

REASONS FOR PERFORMING STUDY: Preputial and penile tumours are more common in horses than in other domestic animals, but no large surveys of male horses with tumours of the external genitalia are available. OBJECTIVE: To present a retrospective analysis of male horses with neoplasms of the external genitalia. METHODS: The penile and preputial tumours of 114 horses were evaluated. Data recorded included age, gelding or stallion and breed; type and site of lesion; involvement of regional lymph nodes; histopathology (including grading of squamous cell carcinoma); and results of radiographic examination of the thorax. RESULTS: Mean age of horses was 19.5 years with no apparent breed predilection. Common presenting clinical signs were irregularities (e.g. the presence of a mass and/or ulceration) on the integument of the penis and prepuce, and purulent or sanguineous discharge from preputial orifice. Squamous cell carcinoma (SCC) was the most prevalent neoplasm followed by papillomas and melanomas. A basal cell carcinoma, neurofibrosarcoma, adenocarcinoma or fibrosarcoma were each found on single horses. Squamous cell carcinomas with poor differentiation had a higher tendency to metastasise than did more differentiated tumours. CONCLUSIONS: Squamous cell carcinoma is the most common urogenital tumour of the male horse and occurs primarily in old horses. Horses with poorly differentiated SCCs tend to have a higher incidence of regional metastases. Pathology of lymph nodes, even when not palpably enlarged, is a valuable diagnostic exercise. Radiology of the thorax to detect lung metastases is of little value.

Penile and preputial squamous cell carcinoma in the horse: a retrospective study of treatment of 77 affected horses.

REASONS FOR PERFORMING STUDY: The most common penile and preputial neoplasm in the horse is the squamous cell carcinoma (SCC), but no large surveys of treatment and effects of the grade of the tumour, based on the degree of differentiation, on outcome of affected horses are available. OBJECTIVES: Analysis of treatment of male horses affected with SCC of the external genitalia and long-term results of treatment. METHODS: Seventy-seven cases of SCC were evaluated. Data recorded included treatment, outcome, post operative histopathology and retrospective tumour grading. RESULTS: Treatments included: cryosurgery, excision, partial phallectomy, partial phallectomy and sheath ablation, and en bloc penile and preputial resection with penile retroversion and removal of inguinal lymph nodes. The incidence of recurrence after partial phallectomy was 25.6% (10/39) and following incomplete removal was 17.9% (7/39). The incidence of recurrence after en bloc resection with retroversion was 12.5% (1/8). In horses with confirmed inguinal lymph node metastasis, the incidence of recurrence was 25.0% (1/4). Poorly differentiated SCCs were more likely to metastasise than well differentiated SCCs, and there was a greater chance that the treatment would be unsuccessful. The success of treatment, complete removal and in preventing recurrence of the tumour, of male horses with SCC of the external genitalia was 55.7%. CONCLUSIONS: Horses that receive only partial phallectomy for treatment for SCC of the external genitalia have a high incidence of recurrence in contrast to horses that receive an en bloc resection. Tumour grading of SCC can help predict prognosis and guide selection of treatment.

Clinical efficacy of intravenous administration of marbofloxacin in a Staphylococcus aureus infection in tissue cages in ponies.

Tissue cages (TC), implanted subcutaneously in the neck in eight ponies, were inoculated with Staphylococcus aureus (S. aureus) to determine the clinical efficacy of marbofloxacin in the treatment of this infection. From 21 h after inoculation, marbofloxacin (6 mg/kg) was administered intravenously (i.v.) once daily for 7 days. Samples of the tissue cage fluid (TCF) were taken to determine marbofloxacin concentrations (days 1, 3 and 7), using high-pressure liquid chromatography, and numbers of viable bacteria [colony forming units (CFU)] (days 1, 3, 7, 14 and 21). Statistical analysis was used to compare CFU before and after treatment. Clinical signs and CFU were used to evaluate the efficacy of treatment. Although, there was a slight decrease in CFU in all TC initially, the infection was not eliminated by marbofloxacin treatment in any of the ponies and abscesses formed. As the MIC (0.25 microg/mL) did not change during treatment and the concentration of marbofloxacin during treatment (mean concentration in TCF was 0.89 microg/mL on day 1, 0.80 microg/mL on day 3 and 2.77 microg/mL on day 7) was above MIC, we consider that the treatment failure might be attributable to the formation of a biofilm by S. aureus. Based on the present results, i.v. administration of marbofloxacin alone is not suitable for the elimination of S. aureus infections from secluded sites.

Clinical efficacy of local administration of ceftiofur in a Staphylococcus aureus infection in tissue cages in ponies.

Ceftiofur concentrations in an infected and uninfected environment were compared and the efficacy of locally administered ceftiofur was evaluated in an experimental infection with Staphylococcus aureus in tissue cages. Eight ponies had tissue cages (TCs) implanted s.c. on each side of the neck. Into one of the cages 150 mg of ceftiofur was administered and fluid samples were taken to determine ceftiofur concentrations. After 1 week the other TC was infected with S. aureus and subsequently treated with 150 mg ceftiofur administered locally into the TC once daily for 21 days. Samples of fluid were taken to determine ceftiofur concentrations and for bacterial counts. Ceftiofur concentrations did not differ significantly in the infected and uninfected environments after single dose of 150 mg of ceftiofur. Concentrations were considerably in excess of the minimum inhibitory concentration (MIC) of the S. aureus strain used. A marked decrease of viable bacteria in tissue cage fluid (TCF) occurred. In five of seven ponies; however, the infection was not eliminated and abscess formation occurred. Therefore, local application of ceftiofur alone is not advisable for infections with S. aureus in secluded sites in horses, but should be used only with adjunctive therapy.

Clinical efficacy of prophylactic administration of trimethoprim/sulfadiazine in a Streptococcus equi subsp. zooepidemicus infection model in ponies.

Tissue chambers, implanted subcutaneously in the neck in six ponies, were inoculated with Streptococcus equi subsp. zooepidemicus in order to determine the clinical efficacy of prophylactic administration of trimethoprim/sulfadiazine (TMP/SDZ) against this infection. The TMP/SDZ treatment consisted of one intravenous (i.v.) injection of 5 mg/kg TMP and 25 mg/kg SDZ and the same dose of TMP/SDZ per os (p.o.), both given 3 h before inoculation. The oral dose was then repeated every 12 h for 5 days. TMP/SDZ concentrations in tissue chamber fluid (TCF) were above 10 times MIC at the moment of inoculation, and they were maintained at this level or higher throughout the duration of treatment. Trimethoprim/sulfadiazine treatment resulted in a marked reduction of viable bacteria in the tissue chamber but did not eliminate the infection, resulting in abscessation from day 19 onwards in all six ponies. This shows that, even when TCF is not yet purulent, TMP/SDZ is unable to eliminate the streptococci. Therefore, TMP/SDZ should not be the antimicrobial treatment of choice in infections in secluded sites in horses.

[A horse with a badly-healing wound? A case report]. / Een paard met een slecht genezende wond? Een case report.

A 14-year-old Haflinger mare was presented with a wound on the right metatarsus which it had sustained 3 years earlier. The wound had never completely healed but had only recently become a problem. Over a period of several months, the wound became larger, produced a lot of exudate, and the horse became lame on the affected limb. Clinical examination and radiographs failed to reveal the cause of the deterioration. Histological evaluation of tissue removed during debridement of the wound revealed squamous cell carcinoma. Because the tumour had already invaded the bone, the prognosis was unfavourable and the horse was euthanised. Necropsy showed the tumour to have metastasised to the right inguinal area and the adjacent mammary gland.

[Rectal tears in the horse: two case reports]. / Rectumbeschadigingen bij het paard: twee casus.

Rectal tears occur incidentally and often have a poor prognosis. Early recognition of the problem and adequate therapy determine the outcome. The history, clinical examination, therapy, and outcome of two horses with grade IIIb and grade IV rectal tears are discussed.

Diagnostic anaesthesia of the equine lower limb: a comparison of lidocaine and lidocaine with epinephrine.

The anaesthetic potency, onset of action, duration of action, and side effects of lidocaine and lidocaine plus epinephrine for proximal metacarpal block of the lateral and medial palmar nerves were determined. Ten horses were used and legs were injected using a cross-over model with three test solutions: 1) solvent and lidocaine (2%) plus epinephrine (SLE); 2) solvent and lidocaine (2%) without epinephrine (SL); and 3) solvent only (S). The contra-lateral leg was injected with saline (placebo; P). In both the SL and SLE groups, the onset of anaesthesia occurred between 5 and 15 minutes after injection. In the SL group the effect was short-lived, lasting 60 minutes, with a maximum effect at 15 minutes. SLE induced long-lasting anaesthesia in most animals moderate or total anesthesia lasted 6 hours, and in two horses more than 9 hours. The maximum effect was achieved at 60 and 90 minutes. Swelling, the main side effect, was seen halfway between the injection site and the fetlock joint. Swelling on day 1 was minimal in the legs injected with P (105.3%) but was progressively greater with S (129%), SL (136%), and SLE (172%). On day 1 and day 2 the differences between SLE and SL, and between SLE and S were significant (P < 0.05). The difference between S and SL was not significant. On day 3, the swelling had subsided and differences were no longer significant.

Adult anopheline ecology and malaria transmission in irrigated areas of South Punjab, Pakistan.

Surface irrigation in the Punjab province of Pakistan has been carried out on a large scale since the development of the Indus Basin Irrigation System in the late 19th century. The objective of our study was to understand how the population dynamics of adult anopheline mosquitoes (Diptera: Culicidae) could be related to malaria transmission in rural areas with intensive irrigation and a history of malaria epidemics. In this paper we present our observations from three villages located along an irrigation canal in South Punjab. The study was carried out from 1 April 1999 to 31 March 2000. Mosquitoes were collected from bedrooms using the pyrethroid spraycatch method and from vegetation and animal sheds using backpack aspirators. Overall, Anopheles subpictus Grassi sensu lato predominated (55.6%), followed by An. stephensi Liston s.l. (41.4%), An. culicifacies Giles s.l. (2.0%), An. pulcherrimus Theobald (1.0%) and An. peditaeniatus Leicester (0.1%). Most mosquitoes (98.8%) were collected from indoor resting-sites whereas collections from potential resting-sites outdoors accounted for only 1.2% of total anopheline densities, confirming the endophilic behaviour of anophelines in Pakistan. Anopheles stephensi, An. culicifacies and An. subpictus populations peaked in August, September and October, respectively. High temperatures and low rainfall negatively affected seasonal abundance in our area. There were interesting differences in anopheline fauna between villages, with An. culicifacies occurring almost exclusively in the village at the head of the irrigation canal, where waterlogged and irrigated fields prevailed. Monthly house-to-house fever surveys showed that malaria transmission remained low with an overall slide positivity rate of 2.4% and all cases were due to infection with Plasmodium vivax. The most plausible explanation for low transmission in our study area seems to be the low density of Pakistan's primary malaria vector, An. culicifacies. The role of other species such as An. stephensi is not clear. Our observations indicate that, in South Punjab, irrigation-related sites support the breeding of anopheline mosquitoes, including the vectors of malaria. As our study was carried out during a year with exceptionally hot and dry climatic conditions, densities and longevity of mosquitoes would probably be higher in other years and could result in more significant malaria transmission than we observed. To assess the overall importance of irrigation-related sites in the epidemiology of malaria in the Punjab, more studies are needed to compare irrigated and non-irrigated areas.