A Novel Monochromatic Light to Detect Target Specimens Within Endoscopic Ultrasonography-guided Fine-needle Aspiration Biopsy Samples.

BACKGROUND/AIM: Endoscopic ultrasonography (EUS)-guided fine-needle aspiration biopsy (EUS-FNB) enhances the diagnostic capabilities of EUS by providing additional pathological samples. However, detecting the target specimens within the collected samples can be challenging. The aim of this study was to determine the optimal wavelength of light for detection of target specimens within EUS-FNB samples in an animal experiment. MATERIALS AND METHODS: EUS-FNB pancreatic tissue samples were collected from a male beagle (weight, 10 kg), and the samples were illuminated with monochromatic light ranging from 430 to 700 nm in 5-nm intervals. The intensities of the target specimen and blood samples were analyzed using the densitometry of the images obtained through irradiation. RESULTS: We found that transmitted monochromatic light of 605 nm most vividly enhanced the contrast between the target specimens and blood in the samples in the impression of appearance. CONCLUSION: Thus, microscopical observations under transmitted light of 605 nm are optimal for target tissue identification within EUS-FNB samples.

Case report: Diagnosis and treatment of pulmonary choristoma in a newborn calf.

A 4-day-old female Holstein calf presented with a large-sized, protruding mass in its back, at birth. Radiography identified the deformed spinous process in the second and third lumbar vertebras, suggesting spina bifida. Ultrasonography of the back mass revealed anechoic bronchi-like structures and large vessels with rich blood flow running parallel within the homogenous echogenic mass's parenchyma. Doppler ultrasonography also revealed pulsatile vessels entering into the deeper side of the transverse process of the lumbar vertebras at the right-sided base of the protruding mass. These imaging results were helpful for surgical planning, in which a large arterial vessel was sutured at the right-sided mass's base, followed by resection of the mass itself. The mass's resection could be carried out according to the planned surgical procedure, though its invasion was too deep to be resected completely. Histopathology for the resected specimens revealed that the mass mainly had lung-tissue-like structures comprised of bronchi-, bronchiole- and alveoli-like structures, and large vessels, allowing the diagnosis of pulmonary choristoma. Doppler ultrasonography could contribute to the differentiation between the bronchi-like tubular structure and the large arterial vessels on the same images, aiding diagnosis of this disease.

One-Step Laparoscopic Abomasopexy for Left Displacements of the Abomasum in Pregnant Cattle: A Retrospective Study.

Although technical descriptions have been published, the applicability of left-flank laparoscopy for the surgical correction of left displacement of the abomasum (LDA) in pregnant cattle has not yet been fully assessed. This study aimed to investigate the postoperative performance of one-step laparoscopic abomasopexy for the correction of LDA during late pregnancy. Fifteen pregnant Holstein cattle with LDA were treated with one-step laparoscopic abomasopexy between days 259 and 274 of gestation. This technique was performed in a standing position. Under endoscopic observation via trocars introduced from the left flank, the animals underwent a surgical procedure to place the bar part of a toggle-pin-suture (TPS) device into the lumen of the abomasum through a long cannula. A suture of a TPS device was secured to the ventral abdominal wall by using a long applicator. No cases experienced postoperative complications, and all had a normal delivery on postoperative day 17.4 ± 4.9. Three of the 15 animals exhibited foot diseases and mastitis after calving. The reproductive performance was recorded during lactation periods, showing a service conception rate of 2.9 ± 1.2 and 110.5 ± 39.1 open days. The one-year survival rate was 60% (9/15). The positive postoperative results demonstrate the benefits of one-step laparoscopic abomasopexy in pregnant bovine cases with LDA.

Efficacy of Abdominal Ultrasonography for Differentiation of Gastrointestinal Diseases in Calves.

This study investigated the clinical efficacy of abdominal ultrasonography for abomasal dilation in three calves, intestinal volvulus in five calves, intussusception in one calf, and internal hernia in one calf. In the abdominal ultrasonograms of the abomasal dilation cases, this disease was commonly characterized by severely extended lumens, including heterogeneously hyperechoic ingesta without intraluminal accumulations of gas. In the animals with intestinal volvulus and intussusception, a to-and-fro flow was observed to be a common ultrasonographic characteristic that led to suspicion of an intestinal obstruction. The use of abdominal ultrasonography for five cases with intestinal volvulus gave no reason to suspect this disease, despite its efficacy in one case, based on an acutely angled narrowing. Although three of five animals with intestinal volvulus had intestinal ruptures, no ultrasonographic evidence could be obtained. When abdominal ultrasonography was used for one case with intussusception, this pathological condition could be strongly suspected, as a "target" sign was observed. This finding supported surgical intervention for this case, followed by treatment with manual reduction, resulting in a favorable outcome. In terms of the differential and definitive diagnosis for various intestinal diseases, abdominal ultrasonography may be poor at providing indicative evidence, but very helpful for confirming intestinal obstruction.

Correlation between Polymerase Chain Reaction Identification of Iron Acquisition Genes and an Iron-Deficient Incubation Test for Klebsiella pneumoniae Isolates from Bovine Mastitis.

We investigated the correlation between the polymerase chain reaction (PCR) identification of six virulence genes associated with siderophore activation and the iron-uptake system (iron-acquisition genes; iucA, entB, fepA, ybtS, psn, and kfu) in mastitis-associated Klebsiella pneumoniae (K. pneumoniae). The growth of 37 K. pneumoniae isolates from the milk of cows with mild mastitis reared on Japanese dairy farms between October 2012 and December 2014 was examined by incubation in an iron-deficient medium. entB-, fepA-, or ybtS-positive isolates grew significantly better than entB-, fepA-, or ybtS-negative isolates after incubating in an iron-deficient medium for three days. Interestingly, the growth of isolates with 0 and ≥4 PCR-positive iron-acquisition genes in the iron-deficient medium were significantly different by day 2, while isolates with 2, 3, and ≥4 PCR-positive iron-acquisition genes grew significantly better than those with no PCR-positive iron-acquisition genes by day 3. Based on the correlation between the results of PCR and iron-deficient incubation tests, iron-deficient incubation for three days can be used to estimate the presence or absence of iron-acquisition genes in mastitis-associated K. pneumoniae.

Diagnostic and Therapeutic Evaluations of Computed Tomography in Three Calves with Unilateral Otitis Media Treated with Ventral Bulla Osteotomy.

This case report describes the clinical utility of computed tomography (CT) in preoperative evaluation of otitis media in three calves, each exhibiting unilateral head tilt and ear droop. Of the three animals examined by CT, right-sided and left-sided involvements of this disease could be diagnosed in two animals and one animal, respectively, as represented by the accumulations of hyperattenuating contents within the extended tympanic bulla. These cases were treated with a ventral bulla osteotomy, which was conducted between the sternothyroid muscle and the omohyoid muscle via an incision made caudally to the area of the thyroid cartilage. The postoperative outcome was satisfactory in one case. However, the remaining two cases showed unsatisfactory outcomes; one calf died on the 10th postoperative day, as it was already seriously ill. The other animal died suddenly during surgery, potentially due to stimulation of the vagus nerve. The use of CT could allow effective visualization of unilateral lesions, proving helpful for the recommendation of surgical intervention. Based on the unfavorable results in two cases, we note that to prevent surgical complications, the clinical applicability of CT such as the creation of severity criteria and development of imaging-assist procedure should be advanced.

Case Report: Ultrasonography and Magnetic Resonance Imaging of Anterior Segment Dysgenesis in a Calf.

This study includes diagnostic efficacy of the antemortem, combined use of ultrasonography and magnetic resonance imaging (MRI) for the diagnosis of anterior segment dysgenesis. A 7-day-old male Holstein calf presented with progressive unilateral exophthalmos associated with enlargement of the right eyeball soon after birth. Ultrasonography of the enlarged right eyeball showed (1) a 2-cm-thick echogenic parenchymal lesion filling the anterior region of the right eyeball, (2) excess accumulation of the anechoic vitreous humor, and (3) absence of the lens structure. Antemortem examination using T2-weighted and fluid-attenuated inversion recovery MRI revealed a thickened, hyperintense anterior lesion and absence of the lens structure. These imaging findings were suggestive of anterior segment dysgenesis. Antemortem imaging showed no abnormalities other than the abnormal structure and size of the right eyeball; therefore, enucleation of the right eye was performed, which allowed intact healing without suppuration. Ocular ultrasonography enhanced the diagnostic accuracy due to the characteristic ultrasonographic findings of a thickened anterior lesion and absence of the lens structure in the eyeball, suggestive of anterior segment dysgenesis.

Lateral abdominal hernia associated with thin abdominal musculature in a calf.

This study aimed to evaluate the clinical utility of ultrasonography in the diagnosis of a newborn calf presenting with extended swelling within its right flank, in addition to its therapeutic planning. Ultrasonograms of the bilateral flanks identified thinning of the external and internal oblique abdominal muscles in whole areas of the abdominal walls. A right lateral abdominal hernia associated with thin abdominal muscular structures was diagnosed ultrasonographically. The right flank abdominal hernia was successfully reconstructed through a modified Mayo mattress suture. This allowed the overlapping of the two very thin structures of the abdominal walls, resulting in the creation of a thicker structure of the right lateral abdominal walls. Reconstruction of the abdominal walls using this method could prevent re-protrusion of the viscera during calf growth.

Detection of squamous cell carcinoma of presumed pancreatic origin and its metastasis in a spotted seal (Phoca largha) using ultrasonography and computed tomography.

A 21-year-old female spotted seal (Phoca largha), with a swollen abdomen, had a five-month history of anorexia and vomiting. Ultrasonography revealed an extended mass with central necrotic foci in the right cranial abdomen. Computed tomography revealed an abdominal mass with a low-density central lumen and a pulmonary nodular lesion. Cytology of an abdominal specimen collected through fine-needle aspiration indicated a malignant tumor with round, atypical cells with large nuclei. Three days after diagnosis, necropsy revealed a 10-cm large, solid, whitish mass in the pancreatic parenchyma and multiple small nodules in the liver, spleen, mesentery, lungs, and mediastinal lymph nodes. Histopathological analysis showed prolific neoplastic cells with marked atypia and occasional keratinization. Immunohistochemistry revealed that the neoplastic cells were positive for cytokeratin AE1/AE3 antibody. Thus, the seal was diagnosed with squamous cell carcinoma, of presumed pancreatic origin, which had metastasized to multiple organs.

Ophthalmic findings in a septic calf with the concurrent exhibition of meningitis and endophthalmitis.

The aim of this study was to evaluate the impacts of ophthalmic findings obtained from both macroscopic examination and ocular ultrasonography when diagnosing bovine endophthalmitis. A newborn crossbreed (Japanese black and Holstein breeds) calf was suspected of visual impairment and central nervous system (CNS) symptoms, such as decreased activity and weak drinking performance. This calf was found to display macroscopic signs, such as clouded lens, convergent strabismus, and horizontal nystagmus, in both eyes. On ocular ultrasonography of both eyes, a V-shaped, thickened, hyperechoic structure was present in the anechoic vitreous humors, indicating retinal detachment. The animal died 4 days after the examination. Sepsis was evident in this case, as Escherichia coli was isolated from multiple organs. The autopsy and histological examination revealed meningitis, encephalitis, and secondary hydrocephalus in the CNS, and endophthalmitis and retinal detachment in both eyes. In this case, the ophthalmic findings did not provide definitive evidence for a diagnosis of endophthalmitis. However, this study indicated that retinal detachment might be an ultrasonographic finding that is suggestive of bovine endophthalmitis.

Genetic Characterization of CTX-M-2-Producing Klebsiella pneumoniae and Klebsiella oxytoca Associated With Bovine Mastitis in Japan.

CTX-M-2-producing Klebsiella oxytoca (K. oxytoca) has not received much attention in animal husbandry compared with Klebsiella pneumoniae (K. pneumoniae), a major reservoir of extended-spectrum ß-lactamase (ESBL) genes. Bacteriological examinations of 1,466 mastitic milk samples between October 2012 and December 2014 were conducted. Ninety-five K. pneumoniae isolates (total prevalence: 6.5%) and 81 K. oxytoca isolates (total prevalence: 5.5%) were obtained. Seventeen K. pneumoniae isolates obtained from 15 animals reared on 11 farms and 9 K. oxytoca isolates obtained from 9 animals reared on the same farm were phenotypically confirmed to be ESBL producers. All nine ESBL-producing K. oxytoca isolates were obtained from one farm between June and November 2013 and related to a significantly (p < 0.05) higher monthly prevalence of mild mastitis (in June, August, September, October, and November 2013). Pulsed-field gel electrophoresis (PFGE) patterns of ESBL-producing K. pneumoniae isolates were distinguished from each other by more than 6-band differences except for two isolates from two animals, whereas all nine K. oxytoca isolates showed an identical PFGE pattern. Transferability of the bla CTX-M-2 gene was found in 14 K. pneumoniae and 9 K. oxytoca isolates by conjugation analysis. Of these isolates, the bla CTX-M-2 gene was detected on plasmids belonging to the incompatibility (Inc) groups P and N derived from five K. pneumoniae and nine K. oxytoca isolates, respectively, although the plasmids from the remaining nine K. pneumoniae were untypeable. All the transconjugants exhibited elevated minimum inhibitory concentrations of ampicillin, cefotaxime, and ceftiofur compared with those in the wild-type, recipient strain. Restriction fragment length polymorphism analysis demonstrated that the IncN plasmids extracted from eight of nine transconjugants, which received resistance against ß-lactams from K. oxytoca, showed an identical DraI digestion pattern. These results suggest that the CTX-M-2-producing K. oxytoca strain with the above-mentioned characteristics may have clonally spread within a farm, whereas the bla CTX-M-2 gene in K. pneumoniae possibly disseminated among the farms through different plasmids. Thus, monitoring of ESBL genes, including the bla CTX-M-2 gene, among causative agents of bacterial mastitis in cows can help to develop relevant treatments and control practices.

Triple nostrils in a calf.

BACKGROUND: Nasal abnormalities are rare in bovines. In humans, nasal deformities are mainly classified as proboscis lateralis or supernumerary nostrils. This report discusses the etiology of triple nostrils in a calf, based on computed tomography, magnetic resonance imaging, and endoscopy. CASE PRESENTATION: A female Holstein calf presented with triple nostrils. The following abnormalities were observed: (1) formation of a small and flat blind-ended middle nostril between the right and left nostrils; (2) presence of a hair-bearing surface on the muzzle; (3) abnormal curvature of the nasal septum, resulting in a narrower right nasal cavity due to transformation of the nasal bones; and (4) formation of a bone-like structure within the nasal septum. These findings were similar to those of supernumerary nostrils in humans. CONCLUSIONS: To the best of our knowledge, this represents the first description of a calf with triple nostrils. The use of imaging modalities is necessary for investigating the etiology of triple nostrils.

Photohyperthermal therapy using liposomally formulated indocyanine green for feline nasal lymphoma: A case report.

Our previous research has focused on the development of a novel cancer therapy by using photohyperthermal therapy (PHT) with indocyanine green (ICG) as an optical sensitizer. ICG-Lipo is a liposomally formulated ICG derivative in which ICG is tagged with an octadeca-alkyl chain to incorporate into liposome bilayers, and contains antitumor drugs such as carboplatin and paclitaxel within the inner membrane space. The present study reported a case of feline nasal lymphoma that was treated with combination therapy of PHT with ICG-Lipo. An antitumour effect was observed, and the patient entered remission. Complications from the radiation treatment included skin burns and bleeding from the irradiated hard palate. Serious side effects related to the drugs were not observed. This report suggested that PHT using ICG-Lipo enabled efficient and safe treatment of lymphoma, and that treatment with a liposomal drug delivery system was enhanced by PHT.

Ultrasonographic changes associated with the healing process in a fracture of the distal phalanx in a cow.

A female Holstein cow presented with lameness in the left hind-limb, having suffered a fracture within the distal phalanx distal to the distal interphalangeal joint. The lesion was visualized as a radiolucent gap on a lateral radiograph and a hypoechoic gap on an ultrasonogram in which the hyperechoic bony line disappeared. Using regular ultrasonographic examinations, the healing of the fracture could be characterized by increased echogenicity inside the gap and the formation of a hyperechoic bony bridge. Ultrasonography can provide earlier evidence compared with that provided by radiography of the healing process of a fractured distal phalanx.

Ultrasonography of sudden swollen tongue in a calf.

BACKGROUND: In cattle, the lingual diseases are primarily diagnosed postmortem by histopathological examination of the affected tongues obtained after the death or during necropsy. In humans, ultrasonography has been used to provide differential diagnoses, and for preoperative or intraoperative planning of glossectomy in various lingual diseases. This is a bovine clinical case report, in which ultrasonography for sudden swelling of the tongue, which was possibly caused by snake bite, was utilized as a preoperative indication to perform a glossectomy. CASE PRESENTATION: An eight-month-old female Japanese black calf presented with sudden swelling of the tongue with well-defined discoloration in the cranial region. A 10-MHz linear probe on a portable-type ultrasound machine (MyLabOne VET, Esaote Co., Genova, Italy) was applied to the ventral surface of the tongues in the affected case, and also in five healthy calves under sedation to observe normal tongues. Ultrasonography of the swollen tongue in this case revealed that the ventral lingual muscular layers were severely thickened compared with those of normal tongues. However, the muscle layers were regularly aligned with the echogenic muscular fibers. This resembled the lingual muscular architectures of normal tongues. Color-flow Doppler ultrasonography revealed that blood flow was weakened in the small peripheral vessels in the spaces between the lingual muscular structures, and was lacking in the deep lingual artery between the apex and base of the tongue. This finding was very different than that of normal tongues, which exhibited weakened or rich blood flows. Based on ultrasonographic findings, this case was treated with glossectomy. After recovery, the calf grew up normally with a normal appetite and rumination, and did not exhibit mouth pain behavior. Histopathologically, hemorrhagic necrotic changes, together with focal formation of fibrin thrombus in the lingual blood vessels in the affected tongue, were noted. CONCLUSIONS: To the best of our knowledge, the present report is the first description of lingual ultrasonography performed in cattle. In this case, ultrasonography enabled visualization of decreased vascularity, which might be associated with hemorrhage or formation of fibrin thrombus in the suddenly swollen tongue presented.

Photodynamic detection of a feline meningioma using 5-aminolaevulinic acid hydrochloride.

CASE SUMMARY: The present study describes the case of a feline meningioma that was detected using 5-aminolaevulinic acid hydrochloride (5-ALA)-induced protoporphyrin IX (PpIX) fluorescence at surgery. An extra-axial mass in the temporoparietal region was observed by MRI. Following craniectomy and durotomy, photodynamic detection (PDD) was performed for detection of the tumour. Intratumour PpIX was detected using fluorescence spectrum evaluation and high-performance liquid chromatography. PDD revealed bright fluorescence of PpIX induced by 5-ALA, facilitating fluorescence-guided resection of the tumour tissue. Postoperative examination demonstrated an intratumour PpIX protein concentration of 16.8 nmol/g, and based on histopathological findings we diagnosed the mass as meningioma. RELEVANCE AND NOVEL INFORMATION: PDD using 5-ALA has been used to identify the surgical margins during resection of primary human brain tumours. Recently, we have reported post-mortem PDD using 5-ALA for a canine glioblastoma. To our knowledge, this technique has not been previously used for the detection and resection of feline brain tumours. Our findings suggest that PDD using 5-ALA is useful for intraoperative fluorescence-guided resection of malignant meningioma in cats.

Diagnostic efficacy of imaging and biopsy methods for peritoneal mesothelioma in a calf.

BACKGROUND: Peritoneal mesothelioma is a rare abdominal disease; that occasionally occurs congenitally in younger calves. Cytologic examination of peritoneal effusion (PE) was utilized to diagnose this disease, and was not diagnostic. Diagnostic accuracy has been elevated by recent use of ultrasonography (US), despite most diagnoses have been obtained post-mortem in slaughter houses or during clinical necropsy. In humans, ante-mortem diagnosis is highly associated with clinical use of computed tomography (CT) and laparoscopy together with imaging-assisted biopsy. The present report evaluates the diagnostic applicability of CT and laparoscopy as well as US via the practical application of these imaging modalities in an affected calf, and compares the cytologic and histologic findings among in PE, and specimens obtained from fine-needle aspiration and core-needle biopsy. In addition, the present results were reviewed in comparison with those of previous bovine and human reports. CASE PRESENTATION: A 58-day-old male Japanese black calf presented first with scrotal swelling, followed by progressive abdominal distention. Abnormalities of the case included: 1) accumulation of anechoic PE inside the swollen scrotum and abdomen; 2) formation of multiple echogenic nodules within the peritoneal membrane based on US images; 3) presence of hyper-dense spots (suspected calcification) along the margins of the nodules; 4) anatomic connections between intra-abdominal nodular lesions and the swollen tunica vaginalis via the inguinal region based on CT images; 5) serosanguineous-colored and less-turbid characteristics of PE; and 6) formation of multiple nodules over all of the serosa of the rumen as well as the peritoneal wall based on laparoscopic views. Fine-needle aspiration and core-needle biopsy were successfully performed under US and laparoscopic observations, respectively. Histology findings of the core-needle biopsy specimen appeared more indicative (characterization of tubular structures comprised of cubical or columnar abnormal mesothelial cell linings) diagnostically of peritoneal mesothelioma than did findings of the fine-needle aspiration specimen. CONCLUSIONS: To the best of our knowledge, this report is the first description of clinical applications of CT and laparoscopy to diagnose peritoneal mesothelioma in a calf. Laparoscopy enhanced the diagnostic accuracy due to clear gross visualization of the intra-abdominal abnormalities and applicability to imaging-guided core-needle biopsy.

Efficacy of 5-Aminolevulinic Acid in Photodynamic Detection and Photodynamic Therapy in Veterinary Medicine.

5-Aminolevulinic acid (5-ALA), a commonly used photosensitizer in photodynamic detection (PDD) and therapy (PDT), is converted in situ to the established photosensitizer protoporphyrin IX (PpIX) via the heme biosynthetic pathway. To extend 5-ALA-PDT application, we evaluated the PpIX fluorescence induced by exogenous 5-ALA in various veterinary tumors and treated canine and feline tumors. 5-ALA-PDD sensitivity and specificity in the whole sample group for dogs and cats combined were 89.5 and 50%, respectively. Notably, some small tumors disappeared upon 5-ALA-PDT. Although single PDT application was not curative, repeated PDT+/-chemotherapy achieved long-term tumor control. We analyzed the relationship between intracellular PpIX concentration and 5-ALA-PDT in vitro cytotoxicity using various primary tumor cells and determined the correlation between intracellular PpIX concentration and 5-ALA transporter and metabolic enzyme mRNA expression levels. 5-ALA-PDT cytotoxicity in vitro correlated with intracellular PpIX concentration in carcinomas. Ferrochelatase mRNA expression levels strongly negatively correlated with PpIX accumulation, representing the first report of a correlation between mRNA expression related to PpIX accumulation and PpIX concentration in canine tumor cells. Our findings suggested that the results of 5-ALA-PDD might be predictive for 5-ALA-PDT therapeutic effects for carcinomas, with 5-ALA-PDT plus chemotherapy potentially increasing the probability of tumor control in veterinary medicine.

Symmetric undivided diplopagus with cardiac malformation in a Japanese wild boar (Sus scrofa leucomystax).

A captured Japanese wild boar (Sus scrofa leucomystax) fetus was dicephalic. The fetus had two heads, but one body from the cranial neck region. Computed tomography imaging revealed that the two crania merged at the occipital bone, and the vertebral bodies between the atlas and the seventh thoracic vertebra were deformed. The fetus was found to have two tongues and laryngopharynges, but its esophagus and trachea were not duplicated. Each head contained a cerebrum and cerebellum, but the brains merged at the obex of the medulla oblongata, and the cervical spinal cord had duplicated ventral clefts. The heart was composed of three atria and four ventricles. This is the first report of a dicephalus with cardiac malformation in a wild boar.

Hair growth-promoting activities of chitosan and surface-deacetylated chitin nanofibers.

In this study, the effects of chitosan and surface-deacetylated chitin nanofibrils (SDACNFs) on hair growth were evaluated. In human follicle dermal papilla cells in vitro, chitosan and SDACNFs were shown to increase cell growth on day 3 after the initiation of treatment, together with an increase in the production of fibroblast growth factor-7 (FGF-7) by these cells on day 3. Furthermore, in an in vivo study in mice, chitosan and SDACNF application promoted hair growth. The number of anagen follicles significantly increased compared with that in the control group, whereas the number of telogen follicles significantly decreased in the chitosan and SDACNF groups. In the chitosan and SDACNFs groups, moreover, the expression levels of FGF-7 and Sonic hedgehog were significantly upregulated in hair follicles. Overall, our results demonstrated that chitosan and SDACNFs promoted hair growth and therefore may have applications as novel therapeutic agents for the treatment of hair loss in patients.