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Károly Vörös Department and Clinic of Internal Medicine, University of Veterinary Medicine Budapest, István u. 2, H-1078, Hungary

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Zsolt Becker Department and Clinic of Internal Medicine, University of Veterinary Medicine Budapest, István u. 2, H-1078, Hungary

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Zoltán Dudás Györki Department and Clinic of Internal Medicine, University of Veterinary Medicine Budapest, István u. 2, H-1078, Hungary

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Bernd Schulze Gronover Department and Clinic of Internal Medicine, University of Veterinary Medicine Budapest, István u. 2, H-1078, Hungary

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Ferenc Szalay Department of Anatomy and Histology, University of Veterinary Medicine Budapest, István u. 2, H-1078, Hungary

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Abstract

Melarsomine is used intramuscularly to destroy adult heartworms when treating canine heartworm disease (HWD). This drug is highly irritative and can elicit local complications. Therefore, melarsomine should be injected into the paralumbar muscles by strictly adhering to the manufacturers' prescriptions. However, it is not known how to determine the optimal location of the needle during the injection process. Ultrasonography (US) of the epaxial (paralumbar) musculature was used as a new method to measure the cross-sectional diameter of the paralumbar musculature, to determine the required location of the injection needle, and to study the local side effects in two dogs with HWD. The macroscopic appearance of the melarsomine solution during injection was demonstrated by video imaging. Melarsomine was not fully gravitating, but its majority was spreading along the thickest fascia of the musculature. Three minutes thereafter, no ultrasound signs of the melarsomine solution were seen, suggesting a full absorption at least ultrasonographically. This procedure was simulated in vitro with methylene blue solution having the same appearance. Removing the injection needle only after 5 min post-injection could prevent undesirable leakage of the drug through the injection channel into the subcutaneous tissue. Ultrasonography can be a useful aid during the treatment of HWD with melarsomine according to this preliminary study.

Abstract

Melarsomine is used intramuscularly to destroy adult heartworms when treating canine heartworm disease (HWD). This drug is highly irritative and can elicit local complications. Therefore, melarsomine should be injected into the paralumbar muscles by strictly adhering to the manufacturers' prescriptions. However, it is not known how to determine the optimal location of the needle during the injection process. Ultrasonography (US) of the epaxial (paralumbar) musculature was used as a new method to measure the cross-sectional diameter of the paralumbar musculature, to determine the required location of the injection needle, and to study the local side effects in two dogs with HWD. The macroscopic appearance of the melarsomine solution during injection was demonstrated by video imaging. Melarsomine was not fully gravitating, but its majority was spreading along the thickest fascia of the musculature. Three minutes thereafter, no ultrasound signs of the melarsomine solution were seen, suggesting a full absorption at least ultrasonographically. This procedure was simulated in vitro with methylene blue solution having the same appearance. Removing the injection needle only after 5 min post-injection could prevent undesirable leakage of the drug through the injection channel into the subcutaneous tissue. Ultrasonography can be a useful aid during the treatment of HWD with melarsomine according to this preliminary study.

Introduction

Heartworm disease (HWD) caused by the filaroid helminth Dirofilaria immitis is a parasitosis widely spread in several continents including Europe (Morchón et al., 2012). The emerging nature of HWD is mainly due to global changing (warming-up) of the climate, by providing even more places for the living conditions of mosquitoes acting as vectors of D. immitis (Fuehrer et al., 2021). Nowadays, HWD has an endemic occurrence also in several Southern and Middle-European countries including Hungary (Genchi et al., 2011; Farkas et al., 2014, 2020). The diagnosis of HWD is based on parasitological laboratory methods, and even subclinically infected dogs need to be treated (Nelson et al., 2020; AHS Guidelines, 2020; Becker et al., 2022). The internationally accepted therapeutic scheme is the three-dose alternate melarsomine treatment regimen recommended by the American Heartworm Society (AHS). This therapeutic protocol consists of the application of macrocyclic lactones against microfilariae as well as against L3 and L4 (larvae), doxycycline for the elimination of the symbiotic Wolbachia bacteria, and melarsomine dihydrochloride (henceforward melarsomine) to kill adult heartworms (HWs). Melarsomine, distributed under the commercial name Immiticide by Boehringer Ingelheim (https://dailymed.nlm.nih.gov/dailymed/fda/fdaDrugXsl.cfm?setid=7b8c0f90-412b-4bd1-9b2b-afd8078e8ecf&type=display) or Diroban by Zoetis Inc. (https://www2.zoetisus.com/content/assets/docs/Petcare/diroban-prescribing-information.pdf), is accepted worldwide and this is the only drug authorised by the FDA in the USA against adult HWs (ESCCAP Guidelines 05, 2012; AHS Guidelines, 2020). Melarsomine has a narrow therapeutic range which should be taken into consideration to avoid potential systemic side effects such as coughing, anorexia, vomiting, diarrhoea or, rarely, neurological alterations. This drug is highly irritative when applied intramuscularly and can often cause – usually temporary – local irritation and pain reaction (https://dailymed.nlm.nih.gov/dailymed/fda/fdaDrugXsl.cfm?setid=7b8c0f90-412b-4bd1-9b2b-afd8078e8ecf&type=display; https://www2.zoetisus.com/content/assets/docs/Petcare/diroban-prescribing-information.pdf). Sometimes, more severe sterile inflammation might occur. In addition to the relatively mild, usually reversible local alterations, more severe complications, including sterile abscess formation, have also been described (Maxwell et al., 2014; Bagi et al., 2017). Two reports have also been published on severe neurological complications affecting the relevant segment of the spinal cord (Hettlich et al., 2003; Moore et al., 2013).

As to the directions of the manufacturers, provided in its label, melarsomine should be injected into the epaxial (more precisely into the paralumbar) musculature, being one of the thickest muscles in dogs with a comprehensive blood supply. The latter is important to provide rapid absorption of the drug (Page, 2008). The highly irritative nature of melarsomine is illustrated in the directions of the manufacturers. As such, different injection needles should be used for suction of the drug from the vial for the injection itself to avoid contamination of the tissues along the outer part of the needle. Alternate lumbar sites should be used during the three-dose melarsomine treatment, and superficial injection or leakage should be avoided (https://dailymed.nlm.nih.gov/dailymed/fda/fdaDrugXsl.cfm?setid=7b8c0f90-412b-4bd1-9b2b-afd8078e8ecf&type=display; https://www2.zoetisus.com/content/assets/docs/Petcare/diroban-prescribing-information.pdf). Proper application with strict adherence to the manufacturers' description can decrease the chance for local complications (Hettlich et al., 2003; Moore et al., 2013). Although these directions are quite detailed, it is not known how to determine the optimal positioning and depth of the injection needle within the paralumbar muscles to minimise local reactions and to avoid more severe complications.

There are few publications on ultrasonography being used for imaging the epaxial (paralumbar) muscles both in human and veterinary medicine (Hides et al., 2008; Stubbs et al., 2010; Haro et al., 2013; Freeman et al., 2017, 2019). Recently, Alaman et al. (2022) have reported on the ultrasound-guided approach to the dorsal aspect of the quadratus lumborum muscle (D-QL) and on evaluation of the spread of methylene blue dye in canine cadavers. However, we did not find reports dealing with the application of ultrasonography in vivo to aid any intramuscular injections in dogs.

Our hypothesis was that ultrasonography can help with the injection technique by providing exact measurement of the cross-sectional diameter of the paralumbar muscles, and it can also be used for the follow-up of the local reactions caused by melarsomine. Our research group has recently reported on the application of moxidectin and ultrasound-aided injection of melarsomine during the American Heartworm Society recommended treatment protocol in 44 Dirofilaria immitis infected dogs (Vörös et al., 2022). Although ultrasonography has been applied in that study for optimising the location of melarsomine injection and following the local effects of this drug, no detailed methodology could be provided because of the limited extent of that article. In this paper, we describe the method of ultrasonographically-aided intramuscular melarsomine injection and present some illustrating cases of this technique.

Materials and methods/Results

In the dogs presented in the study, HWD was confirmed by laboratory methods (Becker et al., 2022) and was treated with the AHS therapeutic protocol (Nelson et al., 2018; AHS Guidelines, 2018). Written owner consent was obtained from each owner for the examinations and treatment as well as for the scientific use of the data gained.

Before the administration of the melarsomine injections, each dog was sedated with butorphanol (Alvegesic® 10 mg/mL inj. ALVERRA and WERFFT GmbH, Vienna, Austria) given deeply intramuscularly into the scapular musculature in a dosage of 0.4 mg/kg.

Dogs were separated in a quiet, darkened room together with their owner to avoid any stressors acting against the sedative effects. The aims of sedation were (1) to immobilise the patients during melarsomine injection and to avoid activity of the lumbar musculature and (2) to utilise the analgesic effect of butorphanol during the post-injection phase. A steady state of sedation was achieved in each dog after 25–30 min. In this phase, the patients remained conscious but became calm and less reactive to their surroundings, but were still able to walk with some trembling.

The dogs were positioned in sternal recumbency on the examination table and pre-injection ultrasonography was performed with a MyLab Gold 40 ultrasound machine (ESAOTE, Italy) equipped with a 6–8 MHz convex abdominal and a 12–18 MHz linear transducer. Ultrasonography of the paralumbar muscles was done as described for the multifidus muscle in humans (Hides et al., 2008). The location of the approximate injection site was chosen between the 3rd and 5th lumbar vertebrae according to the manufacturers' guidance. This was determined by palpating and counting the spinal processes of the lumbar vertebrae, starting just behind the ribcage (i.e., after the last thoracic vertebra). Meanwhile, the paralumbar musculature was also palpated to estimate its most prominent/thickest part. The hair was clipped at this area, copious ultrasound gel was applied, and oblique horizontal planes were produced with the ultrasound probe. These planes were aimed to be perpendicular to the paralumbar muscles, whilst achieving and measuring the largest diameter (thickness) of the musculature. Then, the ultrasound gel was removed carefully and the optimal site for the injection was marked with a felt marker and the area was disinfected with alcohol. The patient was immobilised by the owner and by one assistant. Melarsomine injection was applied as demonstrated in Fig. 1 in dog 1, a Hungarian Vizsla of 27 kg body weight. The needle was inserted approximately into the middle of the muscles based on the pre-injection measurements described above.

Fig. 1.
Fig. 1.

Demonstration of the injection procedure of melarsomine into the previously, ultrasonographically determined intramuscular location in dog 1

Citation: Acta Veterinaria Hungarica 70, 4; 10.1556/004.2022.00034

In Dog 2, a mixed breed of 29.4 kg body weight, the injection needle was removed right after the injection and the injection site was compressed with a gauze pad for 10 min. Meanwhile, the dog was still restrained on the examination table. These measures were intended to avoid the movement of the dog during the absorption of melarsomine from the muscles. Despite these actions, the melarsomine solution was leaking and flowing out from the injection channel to the surface of the skin at the injection puncture site when the needle was pulled out. Then, severe post-injection swelling with moderate pain was observed a few days later. The course of the ultrasonographic alterations of dog 2 is demonstrated in Figs 2–4. This dog had a body condition score (BCS) of 4/5 with a relatively thick subcutaneous tissue layer which might have contributed to the leakage.

Fig. 2.
Fig. 2.

Physiological, oblique-horizontal sonographic image, more or less perpendicular to the paralumbar muscles in dog 2 before melarsomine injection. The echogenic lines represent the intramuscular connective tissue spaces of the musculature (thoracolumbar fascia). One of them is marked with an arrowhead. The thickness (cross-sectional diameter) yields 1.7 cm (between crosses). An 18 MHz linear transducer was used for this image. Legends: lat: lateral, med: medial, cran: cranial, caud: caudal

Citation: Acta Veterinaria Hungarica 70, 4; 10.1556/004.2022.00034

Fig. 3.
Fig. 3.

Oblique-horizontal sonographic image, more or less perpendicular to the paralumbar muscles in dog 2, one day after melarsomine injection. Around the site of the injection the ultrasound structure is homogeneous, this region is more echogenic (arrows). The musculature is apparently thickened due to the local swelling caused by the inflammation as compared to Fig. 2. The cross-sectional diameter yields 3.4 cm (between crosses). An 18 MHz linear transducer was used for this image. Legends: lat: lateral, med: medial, cran: cranial, caud: caudal

Citation: Acta Veterinaria Hungarica 70, 4; 10.1556/004.2022.00034

Fig. 4.
Fig. 4.

Oblique-horizontal sonographic image, more or less perpendicular to the paralumbar muscles in dog 2 shown also on Fig. 3, eight weeks after melarsomine injection and local symptomatic treatment with ice-compress and diclofenac diethylamine ointment. The sonographic structure is nearly normal compared to Fig. 3. The echogenicity of the muscles is only moderately echogenic around the site of the injection and along the intramuscular connective tissue spaces of the musculature (thoracolumbar fascia, arrows). One of them is also marked with arrowhead, the musculature is only mildly thickened as the cross-sectional diameter yields 2.2 cm (between crosses). An 18 MHz linear transducer was used for this image. Legends: lat: lateral, med: medial, cran: cranial, caud: caudal

Citation: Acta Veterinaria Hungarica 70, 4; 10.1556/004.2022.00034

Therefore, the injection technique was modified in the way that the needle was left in place for 5 min after injection of the melarsomine solution when still providing immobilisation of the patient. In dog 3, a Miniature Dachshund dog of 5 kg body weight, we recorded the injection process right through the event (Suppl. Video 1). During this process, we experienced that the drug was not fully gravitating, i.e., flowing to the deepest point of the musculature, towards the vertebrae, but its majority was spreading (more or less) horizontally along the deep thoracolumbar fascia of the musculature (see as an echogenic line on this video recording). Then, 3–5 min after the injection, only minimal ultrasonographic signs of the melarsomine solution were observed, indicating a nearly full absorption at least ultrasonographically (Suppl. Video 2).

These illustrations show that rather severe alterations were found in dog 2, when the needle was removed right after the injection, compared with dog 3 where the needle was left in place for 5 min after the injection. The macroscopic alterations observed and palpated in the paralumbar region, and the macroscopic healing process (decrease in swelling and local sensitivity) occurred in accordance with diminishing of the ultrasound findings.

To simulate the injection process in vitro, 5–10 mL methylene blue solution was injected into the epaxial lumbar musculature of three carcasses under ultrasound guidance as described above. Shortly after the injection, the spot of injection was dissected. A longitudinal distribution of methylene blue was found along with the muscle fibres in the m. longissimus (Fig. 5). The length of distribution varied between 6 and 10 cm in the bodies.

Fig. 5.
Fig. 5.

Distribution of methylene blue injection applied during in vitro ultrasonography of a dog. The scalpel blade indicates the position of the injection. Legends: lat: lateral, med: medial, cran: cranial, caud: caudal

Citation: Acta Veterinaria Hungarica 70, 4; 10.1556/004.2022.00034

Discussion

We found ultrasonography of the paralumbar musculature to be a useful tool for determining the optimal injection site of the highly irritative melarsomine solution. In this way, the required location of the needle pinpoint could be allocated for the injecting process. Based on our preliminary results, by keeping the injection needle in place for about five minutes seems to provide enough time for the absorption of the drug. This can prevent melarsomine leaking into the subcutaneous tissue when the needle is being removed. According to the results of the current work, it can be hypothesised that this technique can minimise or even prevent local complications. By checking the injection site, it is possible to estimate the potential local consequences and the grade of the healing process as well (Vörös et al., 2022).

Regarding analgesia and sedation before and during the injection period, there is no agreement in the literature, and it is not mentioned in the prescriptions of the manufacturers either. In our recent work on the treatment of 44 dogs with heartworm disease (Vörös et al., 2022), we used the synthetic opioid butorphanol, due to its sedative and analgesic effects with minimal cardiovascular sequelae (Hammond et al., 2008; Vin Veterinary Drug Handbook, 2021). Not sedating the patients can lead to reluctance and movement (or even struggling). This can lead to spreading (leaking) of melarsomine close to the nerves or even to the spinal cord of the relevant lumbar region. This was suspected to happen in the case report of Hettlich et al. (2003), resulting in severe neurological complications. Moore et al. (2013) suggested that an inadvertent application of melarsomine into the epidural space or even into the spinal cord could have been responsible for the neurological complications of their case. Other proposed options might include migration of the solution either along fascial planes or along nerve roots into the epidural space (Hettlich et al., 2003). A gradually expanding epidural abscess was suggested as a cause of chronic progressive myelopathy in a dog and this might have been due to the close location of the injection site to the vertebral column (Moore et al., 2013).

Webster et al. (2014) described the functional anatomy of the epaxial musculature in cadavers of sprinting and fighting dogs. However, the comparative anatomy related to the ultrasound appearance of the paralumbar/epaxial musculature is not known. We have demonstrated the distribution of melarsomine in vivo and simulated it in vitro with methylene blue solution with similar results.

There are obvious limitations of the present work. This is only a description and illustration of the technique without prospective follow-up of clinical cases. The rapid absorption of melarsomine demonstrated in the present study is mentioned but not detailed in the labels of the manufacturers (https://dailymed.nlm.nih.gov/dailymed/fda/fdaDrugXsl.cfm?setid=7b8c0f90-412b-4bd1-9b2b-afd8078e8ecf&type=display; https://www2.zoetisus.com/content/assets/docs/Petcare/diroban-prescribing-information.pdf). Melarsomine is absorbed quickly, and it reaches its maximum concentration in the blood in about 11 min (Page, 2008). However, we did not find literary information on the full post-injection absorption (i.e., disappearance) of this drug from the musculature. The precise determination of melarsomine absorption could be made only by high-performance liquid chromatography (HPLC) of muscular biopsy samples taken from the places of the injection. This is not an option in clinical cases and would need experimental studies. In a recent research, we have successfully applied our currently reported technique in 44 dogs with HWD (Vörös et al., 2022). Those patients were treated according to the AHS recommendation which was expanded with some new therapeutic measures. Melarsomine was injected intramuscularly as described in the present article and post-therapeutic local complications were detected and followed by physical examination as well as by ultrasonography of the paralumbar muscles (Vörös et al., 2022). That publication is partially based on the present preliminary study which has been cited in that paper as ‘under review’. Therefore, the current manuscript can be considered as a preceding preliminary work.

Ethical statement

The first author, Károly VÖRÖS is a member of the Editorial Board.

Acknowledgements

The authors express their thanks to Prof. Róbert Farkas, DVM, PhD, DSc, as well as to Mrs Mónika Gyurkovszky (Department of Parasitology and Zoology of the University of Veterinary Medicine Budapest, UVMB) for performing the parasitological laboratory examinations. This research was funded by the KK 69P02RM06 research project (2017) and by the NKB project (2018), both provided by the UVMB. The study was also supported by the Doctoral School of the UVMB as part of the doctoral thesis of ZB.

Supplementary material

Supplementary video 1. YouTube link: https://www.youtube.com/watch?v=4QlM34AFM7o. Video image of dog 3 during insertion of the needle and distribution of melarsomine within the lumbar musculature (see text for explanation). A 6 MHz convex abdominal transducer was used for this image to allow simultaneous needle insertion and video recording. The resolution of this transducer is lower, compared to the 18 MHz linear transducer applied for Figs 2–4.

Supplementary video 2. YouTube link https://www.youtube.com/watch?v=JSXVDwGX9Ns. Video image of dog 3 after five minutes of melarsomine injection into the lumbar musculature (see text for explanation). A 6 MHz convex abdominal transducer was used for this image to allow simultaneous needle insertion and video recording. The resolution of this transducer is lower, compared to the 18 MHz linear transducer applied for Figs 2–4.

Supplementary data to this article can be found online at https://doi.org/10.1556/004.2022.00034.

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Senior editors

Editor-in-Chief: Ferenc BASKA

Editorial assistant: Szilvia PÁLINKÁS

 

Editorial Board

  • Mária BENKŐ (Acta Veterinaria Hungarica, Budapest, Hungary)
  • Gábor BODÓ (University of Veterinary Medicine, Budapest, Hungary)
  • Béla DÉNES (University of Veterinary Medicine, Budapest Hungary)
  • Edit ESZTERBAUER (Veterinary Medical Research Institute, Budapest, Hungary)
  • Hedvig FÉBEL (University of Veterinary Medicine, Budapest, Hungary)
  • László FODOR (University of Veterinary Medicine, Budapest, Hungary)
  • János GÁL (University of Veterinary Medicine, Budapest, Hungary)
  • Balázs HARRACH (Veterinary Medical Research Institute, Budapest, Hungary)
  • Peter MASSÁNYI (Slovak University of Agriculture in Nitra, Nitra, Slovak Republic)
  • Béla NAGY (Veterinary Medical Research Institute, Budapest, Hungary)
  • Tibor NÉMETH (University of Veterinary Medicine, Budapest, Hungary)
  • Zsuzsanna NEOGRÁDY (University of Veterinary Medicine, Budapest, Hungary)
  • Dušan PALIĆ (Ludwig Maximilian University, Munich, Germany)
  • Alessandra PELAGALLI (University of Naples Federico II, Naples, Italy)
  • Kurt PFISTER (Ludwig-Maximilians-University of Munich, Munich, Germany)
  • László SOLTI (University of Veterinary Medicine, Budapest, Hungary)
  • József SZABÓ (University of Veterinary Medicine, Budapest, Hungary)
  • Péter VAJDOVICH (University of Veterinary Medicine, Budapest, Hungary)
  • János VARGA (University of Veterinary Medicine, Budapest, Hungary)
  • Štefan VILČEK (University of Veterinary Medicine in Kosice, Kosice, Slovak Republic)
  • Károly VÖRÖS (University of Veterinary Medicine, Budapest, Hungary)
  • Herbert WEISSENBÖCK (University of Veterinary Medicine, Vienna, Austria)
  • Attila ZSARNOVSZKY (Szent István University, Gödöllő, Hungary)

ACTA VETERINARIA HUNGARICA
Institute for Veterinary Medical Research
Centre for Agricultural Research
Hungarian Academy of Sciences
P.O. Box 18, H-1581 Budapest, Hungary
Phone: (36 1) 287 7073 (ed.-in-chief) or (36 1) 467 4081 (editor)

E-mail: acta.veterinaria@univet.hu (ed.-in-chief)

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2023  
Web of Science  
Journal Impact Factor 0.7
Rank by Impact Factor Q3 (Veterinary Sciences)
Journal Citation Indicator 0.4
Scopus  
CiteScore 1.8
CiteScore rank Q2 (General Veterinary)
SNIP 0.39
Scimago  
SJR index 0.258
SJR Q rank Q3

Acta Veterinaria Hungarica
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Acta Veterinaria Hungarica
Language English
Size A4
Year of
Foundation
1951
Volumes
per Year
1
Issues
per Year
4
Founder Magyar Tudományos Akadémia
Founder's
Address
H-1051 Budapest, Hungary, Széchenyi István tér 9.
Publisher Akadémiai Kiadó
Publisher's
Address
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Responsible
Publisher
Chief Executive Officer, Akadémiai Kiadó
ISSN 0236-6290 (Print)
ISSN 1588-2705 (Online)

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