Detection of single-nucleotide polymorphisms in Plasmodium falciparum by PCR primer extension and lateral flow immunoassay.

The resistance of Plasmodium falciparum to some antimalarial drugs is linked to single-nucleotide polymorphisms (SNPs). Currently, there are no methods for the identification of resistant parasites that are sufficiently simple, cheap, and fast enough to be performed at point-of-care, i.e., in local hospitals where drugs are prescribed. Primer extension methods (PEXT) were developed to identify 4 SNPs in P. falciparum positioned at amino acids 86, 184, and 1246 of the P. falciparum multidrug resistance 1 gene (pfmdr1) and amino acid 76 of the chloroquine resistance transporter gene (pfcrt). The PEXT products were visualized by a nucleic acid lateral flow immunoassay (NALFIA) with carbon nanoparticles as the detection labels. PCR-PEXT-NALFIAs showed good correlation to the reference methods, quantitative PCR (qPCR) or direct amplicon sequence analysis, in an initial open-label evaluation with 17 field samples. The tests were further evaluated in a blind study design in a set of 150 patient isolates. High specificities of 98 to 100% were found for all 4 PCR-PEXT genotyping assays. The sensitivities ranged from 75% to 100% when all PEXT-positive tests were considered. A number of samples with a low parasite density were successfully characterized by the reference methods but failed to generate a result in the PCR-PEXT-NALFIA, particularly those samples with microscopy-negative subpatent infections. This proof-of principle study validates the use of PCR-PEXT-NALFIA for the detection of resistance-associated mutations in P. falciparum, particularly for microscopy-positive infections. Although it requires a standard thermal cycler, the procedure is cheap and rapid and thus a potentially valuable tool for point-of-care detection in developing countries.

Diagnostic findings in sinonasal aspergillosis in dogs in the United Kingdom: 475 cases (2011-2021).

OBJECTIVES: To describe the diagnostic tests used and their comparative performance in dogs diagnosed with sinonasal aspergillosis in the United Kingdom. A secondary objective was to describe the signalment, clinical findings and common clinicopathologic abnormalities in sinonasal aspergillosis. MATERIALS AND METHODS: A multi-centre retrospective survey was performed involving 23 referral centres in the United Kingdom to identify dogs diagnosed with sinonasal aspergillosis from January 2011 to December 2021. Dogs were included if fungal plaques were seen during rhinoscopy or if ancillary testing (via histopathology, culture, cytology, serology or PCR) was positive and other differential diagnoses were excluded. RESULTS: A total of 662 cases were entered into the database across the 23 referral centres. Four hundred and seventy-five cases met the study inclusion criteria. Of these, 419 dogs had fungal plaques and compatible clinical signs. Fungal plaques were not seen in 56 dogs with turbinate destruction that had compatible clinical signs and a positive ancillary test result. Ancillary diagnostics were performed in 312 of 419 (74%) dogs with observed fungal plaques permitting calculation of sensitivity of cytology as 67%, fungal culture 59%, histopathology 47% and PCR 71%. CLINICAL SIGNIFICANCE: The sensitivities of ancillary diagnostics in this study were lower than previously reported challenging the clinical utility of such tests in sinonasal aspergillosis. Treatment and management decisions should be based on a combination of diagnostics including imaging findings, visual inspection, and ancillary testing, rather than ancillary tests alone.

Short- and long-term outcomes of polyethylene band attenuation of congenital extrahepatic portosystemic shunts in dogs: 60 cases (2010-2020).

OBJECTIVES: To report the short- and long-term outcomes following attenuation of congenital extrahepatic portosystemic shunts in dogs using a novel polyethylene band. MATERIALS AND METHODS: Records were retrospectively reviewed for dogs that underwent congenital extrahepatic portosystemic shunt attenuation by a polyethylene banding technique, at a single institution between 2010 and 2020. Short-term outcome data were collected from peri-operative clinical records with follow-up examinations, scheduled at 6 and 18 weeks post-operatively, and post-operative imaging when performed. Long-term follow-up was collected by validated owner questionnaire, telephone interview or medical records. Long-term outcomes were categorised by "excellent", "good" or "poor". RESULTS: Sixty dogs were included. Post-operative complications occurred in 10 of 60 dogs (16.7%), four major and six minor, with a peri-operative mortality of 6.7%. Persistent shunting was identified in nine of 53 dogs (17%) available for follow-up examination and four dogs underwent a revision surgery. Long-term follow-up was available for 44 dogs at a median of 75 months post-operatively (range 7 to 128). Long-term outcomes were "excellent" (26) or "good" (8) in 81.8% of dogs and "poor" (8) in 18.2%. At the time of follow-up, 30 of 44 (68.2%) dogs were not receiving any medical treatment and 27 of 28 (96.4%) questionnaire respondents were satisfied with the response to surgery. CLINICAL SIGNIFICANCE: Polyethylene band attenuation of congenital extrahepatic portosystemic shunts provides comparable outcomes to cellophane. The material used in this study is widely available and consistent while being pre-sterilised and pre-folded makes it easy to use.

Congenital pleuroperitoneal hernia presenting as gastrothorax in five cavalier King Charles spaniel dogs.

Five cavalier King Charles spaniels were examined for acute onset of respiratory distress. Thoracic radiographs demonstrated diaphragmatic hernia and tension gastrothorax, visible as a distended stomach occupying the left caudal thoracic cavity. Exploratory midline coeliotomy confirmed congenital pleuroperitoneal diaphragmatic hernia with herniation and dilatation of the stomach. The hernia configuration was consistent in all cases, with a defect affecting the left diaphragmatic crus. Congenital pleuroperitoneal diaphragmatic hernia is a rare condition caused by a defect in the dorsolateral diaphragm. Defects of the left crus of the diaphragm could result in the herniation of the stomach into the thoracic cavity with possible subsequent tension gastrothorax. Cavalier King Charles spaniels may have a predisposition to this condition. Tension gastrothorax is an acute life-threatening consequence of gastric herniation through a diaphragmatic defect that must be promptly recognised and surgically treated.

Study of how frequently surgeons' gloves are perforated during operations.

Surgical gloves were collected after 231 first opinion and referral surgical procedures performed at the University of Liverpool Small Animal Hospital. They were tested for defects by the water load test and the site of any defects was recorded; 10.2 per cent of the gloves had defects, and at least one glove became defective during 51 (22.1 per cent) of the surgical procedures. Significantly more defects occurred during orthopaedic procedures and the likelihood of a defect occurring increased with the duration of the surgical procedure. The majority (78 per cent) of the defects occurred in the gloves worn on the non-dominant hand and 64 per cent of them were in the index finger. The surgeon or assistant was aware of a glove being punctured in only 3.8 per cent of cases.

Prospective evaluation of postoperative pain in cats undergoing ovariohysterectomy by a midline or flank approach.

Twenty entire female cats were randomly assigned to two groups of 10; the cats in one group underwent ovariohysterectomy by a midline approach and the cats in the other group by a flank approach. Cats were assessed for signs of pain and scores were assigned pre- and postoperatively. There was a tendency for the cats neutered by a flank approach to be in more pain postoperatively (P=0.05). The final pain score for cats in either group was equal to or lower than their baseline score.

Pyloric localisation in 57 dogs of breeds susceptible to gastric dilatation-volvulus in the UK using computed tomography.

OBJECTIVE: Describe the location of the pylorus using CT in dog breeds susceptible to gastric dilatation-volvulus in the UK. METHODS: Descriptive anatomical study. Abdominal CT scans of 57 client-owned dogs were reviewed to assess pyloric position relative to the 9th, 10th, 11th and 13th ribs and 2 and 3 cm caudal to the 13th rib at the 8, 9 and 10 o'clock positions. The angle of the pylorus from the centre of the abdominal cavity relative to the sagittal plane was also determined. RESULTS: In 88 per cent of cases, the pylorus was located in the right cranioventral abdomen with 63 per cent positioned at the 9-10 o'clock position. The overall distance between the pylorus and right abdominal wall (RAW) at the 13th rib 10 o'clock position was equivalent to 29 per cent of ventral abdominal length, significantly greater than the median overall distance of ∼14 per cent of ventral abdominal length between the pylorus and RAW at the 9th or 10th rib 10 o'clock position (P<0.0001). CLINICAL SIGNIFICANCE: Common gastropexy locations may result in considerable displacement of the pylorus relative to its natural anatomic location. Further case-control studies are required to assess the clinical significance of this finding.

Clinical findings and treatment outcomes for cats diagnosed with patent ductus arteriosus in the UK: a retrospective study of 19 cases (2004-2012).

Patent ductus arteriosus (PDA) is infrequently reported in cats and represents between 1-7.3 per cent of left to right shunting cardiac congenital anomalies. The objective of this study was to report the presenting complaints, clinical examination findings, diagnostic findings, treatment outcomes and survival times in cats diagnosed with a PDA in the UK. Medical records from three major UK referral centres were searched for cats that were diagnosed with PDA from January 2004 to December 2012. Data obtained for analysis included: signalment, clinical examination findings including murmur characteristics, diagnostic imaging findings, treatment outcomes and survival times. Nineteen cats were included in the analysis. The most common reason for referral was investigation of an incidentally detected heart murmur without clinical signs (13/19; 68 per cent). Pulmonary arterial hypertension (PAH) was diagnosed in seven (37 per cent) cats and those cats with PAH were significantly more likely to present with signs of disease (P=0.004). Median survival time in cats that were diagnosed with PDA and died due to cardiac causes was 898 days (interquartile range 459-1011 days). The median survival time of those cats that had an additional congenital anomaly was significantly shorter to those cats without a congenital anomaly (P=0.008).

Canine pancreatic islet cell tumours secreting insulin-like growth factor type 2: a rare entity.

Insulin-like growth factor type II (IGF-II) is the main cause of non-islet cell tumour hypoglycaemia (NICTH) and insulin is thought to be the only factor causing hypoglycaemia in insulinomas. However, two case reports of pancreatic neuroendocrine tumours (PNETs) producing IGF-II have been previously published: a human and a canine patient. In this study, we investigated clinical, histopathological, immunohistochemical and ultrastructural features, and biological behaviour of canine pancreatic IGF-II-omas, a subgroup of PNETs that has not been previously characterized. Case records of 58 dogs with confirmed PNETs and hypoglycaemia were reviewed: six patients were affected by IGF-II-omas. Surgery was performed in all cases and two dogs had metastases. Four patients remained alive and in remission at 370, 440, 560 and 890 days post-diagnosis; two died of non-tumour-related causes. IGF-II-omas can be differentiated from insulinomas through hypoinsulinaemia, IGF-II positive and insulin negative immunostaining. The prevalence of this neoplasia is low, accounting for just 6% of PNETs.

Complications observed during and after ovariohysterectomy of 142 bitches at a veterinary teaching hospital.

The clinical, anaesthetic and surgical records of 142 bitches admitted into the primary care clinic at the Small Animal Hospital, University of Liverpool, between January 2002 and August 2004 for routine ovariohysterectomy by final-year students were analysed for complications during and after the surgery. The time taken to perform the surgical procedure was recorded, to allow assessment of the time involved in one-to-one surgical teaching during live-recovery surgeries of client-owned pets. The types of complications were similar to those reported in earlier studies: nine bitches suffered haemorrhage of the ovarian arteries during surgery, and there were four cases of haemorrhage after surgery, 12 cases of wound inflammation and four other types of complication. The rates of intraoperative, postoperative and total complications were 6.3, 14.1 and 20.6 per cent, respectively.

Thrombo-ischaemic pinnal necrosis associated with fenbendazole treatment in a dog.

An 11-week-old, female West Highland white terrier was presented with necrosis of the distal third of both pinnae. Haematology, biochemistry and urinalysis, Coombs test, antinuclear antibody and cold autoagglutinin antibody tests were normal. A drug reaction to fenbendazole was diagnosed. The necrotic ear tips were surgically removed. Histopathology revealed extensive coagulative necrosis of the epidermis and superficial to mid-dermis, a moderate interstitial neutrophilic infiltrate and complete thrombotic occlusion and necrosis of blood vessels. There was also endothelial cell activation and proliferation with endothelial cell cushions protruding into the vascular lumen. Immunohistochemistry for factor VIII-related antigen confirmed endothelial cell involvement. This case represents an unusual, drug-induced, thrombo-ischaemic necrosis of the pinnae. It is also, to the authors' knowledge, the first report of fenbendazole sensitivity in a dog. The histopathology is similar to previous cases of proliferative thrombovascular pinnal necrosis, suggesting that drug reactions should be considered in this condition.

Diagnostic accuracy of indium-111 platelet scintigraphy in identifying left ventricular thrombi.

This study defines the optimum imaging time window after injection of labeled platelet suspension for detection on left ventricular (LV) thrombi, identifies the most useful imaging views, and determines the reproducibility of this technique. A total of 662 images obtained from 64 patients were analyzed retrospectively on 2 separate occasions by 3 observers blinded as to patient identity, view (right anterior oblique, anterior, left anterior oblique, and left lateral), and time after injection of the platelet suspension that the images were obtained (0 to 2, 3 to 4, and 5 to 6 days). Images were categorized as either positive or negative. In every case surgical or autopsy verification of the presence or absence of LV thrombus was possible. The best combination of sensitivity, specificity, and diagnostic accuracy was found in the 3- to 4-day period in the left anterior oblique view and was 54 +/- 5% (mean +/- standard deviation), 98 +/- 1%, and 85 +/- 2%, respectively. Sensitivity, specificity, and diagnostic accuracy were not enhanced by adding additional views (right anterior oblique, left lateral, and anterior) to the left anterior oblique view in the 3- to 4-day time period. However, using multiple views, localization of thrombi to the left ventricle was facilitated. In a second retrospective analysis, a comparison of day 0 with day 3 to 4 images enhanced sensitivity and accuracy to 65 (p less than 0.001) and 90% (not significant), respectively. Specificity was unchanged at 99%. Mean intra- and interobserver agreement was 91 and 88%, respectively. Thus, (1) indium-111 platelet scintigraphy is a reproducible and specific technique for identifying LV thrombus, and (2) we advise imaging on day 0 and again 3 to 4 days after injection of the platelet suspension in right anterior oblique, left anterior oblique, left lateral, and anterior views to maximize accuracy and to facilitate localization of LV thrombus.

Diterpenoids from Humirianthera ampla.

Two diterpenoids, humirianthol and acrenol, as well as the known annonalide, were isolated from Humirianthera ampla. Humirianthol and acrenol were determined by 1D and 2D NMR spectroscopic techniques to be 3 beta,20:14 beta,16-diepoxy-3 alpha, 15 alpha-dihydroxy-7-pimaren-19,6 beta-olide and 3 beta,20-epoxy-3 alpha,15,16-trihydroxy-7-pimaren-19,6 beta-olide, respectively.

The impact of dedicated physician staffing on patient flow and quality assurance parameters in an Air Force emergency department.

A retrospective audit of 17,028 emergency charts at USAF Medical Center, Scott was performed over two time periods to compare patient waiting times and selected quality assurance parameters with two methods of physician staffing. Phase 1 consisted of 4 months when the Emergency Department (ED) was manned with five physicians assigned only to that department. Non-departmental physicians supplemented the full-time staff. Phase 2 was the corresponding 4 months the following year when the ED was staffed with six emergency physicians, and non-emergency physician coverage was minimal. The difference in daily census between the two periods of time was not statistically significant. The time required to be seen by a physician decreased from an average of 25.6 minutes per patient in phase 1 to 13.7 minutes per patient in phase 2. Time to disposition also decreased from 71.9 minutes per patient in phase 1 to 59.5 minutes per patient in phase 2. In the second phase the number of "positive" x-ray findings increased, while the number of incomplete charts and patients who left without being seen by a physician diminished.

A nasal dermoid sinus in an English bull terrier.

A five-year-old, entire female English bull terrier was presented with a six-week history of intermittent facial swelling and nasal pain, following an incident of nasal trauma. A small opening was present at the mucocutaneous junction on the dorsal nasal planum. Examination under general anaesthesia allowed catheterisation of this opening and confirmed the presence of a tract passing caudally. Plain radiographic examination of the region was unremarkable. Positive contrast sinography demonstrated contrast material filling a midline tract that passed caudally and subcutaneously towards the nasal bone. This tract was surgically excised. Histopathological examination of the excised tissue, together with the clinical findings, led to the diagnosis of nasal dermoid sinus.

Frontal sinus depth at four landmarks in breeds of dog typically affected by sinonasal aspergillosis.

The objective of this study was to assess whether the frontal sinuses in dogs with aspergillosis and of breeds typically affected by this condition were deeper at a more caudal location. CT scans of the head performed at the Small Animal Teaching Hospital, University of Liverpool, between April 2007 and March 2009 for dogs diagnosed with aspergillosis (group 1) and unaffected dogs of similar breeds (group 2) were selected for study. Sinus depth was measured at four standardised locations from reconstructed images of these CT scans. Data were compared for differences in sinus depth between groups and between landmarks. No significant difference was found between measurements within individual dogs or for each of the various landmarks between groups. Difference in depth of the sinuses between landmarks was significant (P<0.001). Sinus depth was significantly greater at the more caudal landmarks and was shallowest at the previously recommended landmark for sinus entry. In 54 per cent of dogs, the frontal sinus depth measured less than or equal to 2 cm at one or more of the landmarks. Sinus entry at the deepest point will reduce the risk of accidentally damaging underlying structures. This may be approximately 1 cm caudal, in breeds of dog that typically develop aspergillosis, to a previously suggested landmark.

Retained surgical swabs in 13 dogs.

To report the signalment, history, clinical signs, diagnostic and surgical findings and outcome in dogs and cats with retained swabs, medical records of dogs and cats that were referred to four referral centres were reviewed. Cases in which a retained surgical swab was identified during surgery at the referral hospital were selected for further study. The signalment, history, clinical signs, diagnostic and surgical findings and outcome in these cases were reviewed. Thirteen dogs with retained surgical swabs were included in the study. Bodyweight ranged from 9 to 45 kg. The initial surgery was a 'non-routine' abdominal procedure in seven dogs, ovariohysterectomy in five dogs and perineal hernia repair in one dog. Time from initial surgery to presentation at the referral centre ranged from four days to seven years. Treatment involved major surgery in eight of the 13 dogs. Outcome was excellent in 12 of 13 cases, one dog was euthanased.

Penile amputation and scrotal urethrostomy in 18 dogs.

The objective of this study was to report the signalment, indications for surgery, postoperative complications and outcome in dogs undergoing penile amputation and scrotal urethrostomy. Medical records of three surgical referral facilities were reviewed for dogs undergoing penile amputation and scrotal urethrostomy between January 2003 and July 2010. Data collected included signalment, presenting signs, indication for penile amputation, surgical technique, postoperative complications and long-term outcome. Eighteen dogs were included in the study. Indications for surgery were treatment of neoplasia (n=6), external or unknown penile trauma (n=4), penile trauma or necrosis associated with urethral obstruction with calculi (n=3), priapism (n=4) and balanoposthitis (n=1). All dogs suffered mild postoperative haemorrhage (posturination and/or spontaneous) from the urethrostomy stoma for up to 21 days (mean 5.5 days). Four dogs had minor complications recorded at suture removal (minor dehiscence (n=1), mild bruising and swelling around the urethrostomy site and mild haemorrhage at suture removal (n=2), and granulation at the edge of stoma (n=1)). One dog had a major complication (wound dehiscence and subsequent stricture of the stoma). Long-term outcome was excellent in all dogs with non-neoplastic disease. Local tumour recurrence and/or metastatic disease occurred within five to 12 months of surgery in two dogs undergoing penile amputation for the treatment of neoplasia. Both dogs were euthanased.

The effect of boric acid on bacterial culture of canine and feline urine.

OBJECTIVE: To identify the optimal method of submission of canine and feline urine for bacterial culture. METHODS: Cystocentesis samples from 250 animals (200 dogs, 50 cats) suspected of having urinary tract infections were collected. The reference aliquot, without preservative, was processed on site within 2 hours. Two further aliquots (one without preservative, one with boric acid) were stored at room temperature for up to 7 hours and then posted by guaranteed next day delivery to a commercial laboratory for analysis. RESULTS: Forty-seven of the samples were positive on culture in the reference test. There was no significant difference between reference test results and those of samples posted without preservative (P=0·39), but samples posted in boric acid were significantly less likely to give a positive result (P=0·01). Samples posted without preservative had a sensitivity of 82% and a specificity of 98%; for boric acid, sensitivity was 73% and specificity 99%. CLINICAL SIGNIFICANCE: Postal urine samples should be submitted to the laboratory in a plain sterile tube.

Mast cell tumour and cutaneous histiocytoma excision wound healing in general practice.

OBJECTIVE: To assess whether wounds from incomplete mast cell tumour excisions are at greater risk of healing complications than wounds from complete excisions, or cutaneous histiocytomas. METHODS: Mast cell tumours and cutaneous histiocytomas submitted to Nationwide Laboratories between November 1, 2007 and April 30, 2008 were selected. Questionnaires were sent to submitting veterinarians requesting details of tumour characteristics, clinical approach to the tumour and wound healing. RESULTS: Three hundred and eighty-six mast cell tumours and 524 cutaneous histiocytomas were identified. One hundred and eighty-five mast cell tumours and 244 cutaneous histiocytomas questionnaires were returned (47% response). Wound complications arose in 20% of mast cell tumours and 21% of cutaneous histiocytomas. Multivariable analysis confirmed that larger tumours, tumours on the feet and a soft/"baggy" appearance, were significantly associated with a greater frequency of problems, leading to delayed wound healing and dehiscence. CLINICAL SIGNIFICANCE: Incomplete mast cell tumour excision does not lead to greater risk of wound complications. Mast cell tumour surgical wounds have a similar rate of wound complications as cutaneous histiocytoma wounds.