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Tetanus is a bacterial infection of the nervous system caused by neurotoxins produced by Clostridium tetani. Cephalic tetanus is a rare, localized form of the disease that is often under-recognized and misdiagnosed. Patients typically present with cranial neuropathies, trismus, and stiffness localized to the head and neck. Early recognition of the disease is crucial, as it is easily treatable and can prevent progression to the more severe generalized form. In this report, we describe a patient who developed progressive neurological symptoms after sustaining a forehead laceration from a fall. Over the course of three weeks, he developed facial (VII) nerve palsy, ptosis, followed by trismus, and later orthopnea, which are uncommon manifestations of the disease. This case underscores the importance of early recognition, treatment, and prevention of this neglected condition, which is not commonly observed in well-developed healthcare systems where immunization is universally available. [ABSTRACT FROM AUTHOR]

Tetanus neurotoxins (TeNT) and botulinum neurotoxins (BoNTs) are closely related ~150 kDa protein toxins that together comprise the group of clostridial neurotoxins (CNTs) expressed by various species of Clostridia. While TeNT is expressed as a single polypeptide, BoNTs are always produced alongside multiple non-toxic proteins that form a stabilizing complex with BoNT and are encoded in a conserved toxin gene cluster. It is unknown how tent evolved without a similar gene cluster and why complex-free TeNT is secreted as a stable and soluble protein by C. tetani, whereas complexing proteins appear to be essential for BoNT stability in culture supernatants of C. botulinum. To assess whether the stability of TeNT is due to an innate property of the toxin or is a result of C. tetani's intra- and extra-cellular environment, both TeNT and complex-free BoNT/A1ERY were expressed recombinantly in atoxic C. tetani and analyzed for expression and stability. The strong clostridial ferredoxin (fdx) promotor resulted in the expression of recombinant TeNT at greater levels and earlier time points than endogenously produced TeNT. Recombinant BoNT/A1ERY was similarly expressed by atoxic C. tetani, although partial degradation was observed. The rBoNT/A1ERY produced in C. tetani was also partially proteolytically processed to the dichain form. Investigations of bacterial growth media and pH conditions found that the stability of rTeNT and rBoNT/A1ERY in spent media of C. tetani or C. botulinum was affected by growth media but not by pH. These data indicate that the distinct metabolism of C. tetani or C. botulinum under various growth conditions is a primary factor in creating a more or less favorable environment for complex-free CNT stability. [ABSTRACT FROM AUTHOR]

Background: Parkinson's disease (PD) is a neurodegenerative disorder that progressively damages the autonomic and central nervous systems, leading to hallmark symptoms such as resting tremor, bradykinesia, and rigidity. Despite extensive research, the underlying cause of PD remains unclear, and current treatments are unable to halt the progression of the disease. In this retrospective study, based on historical electronic health records (EHR) from a national health provider covering the period from 2003 to 2023, we investigated the impact of vaccination and medication purchases on PD occurrence and severity. Methods: Using a case–control design, we compared the vaccination histories of 1446 PD patients with 7230 matched controls to assess the association between vaccination and PD onset. Additionally, we explored statistical associations between vaccination, medication purchases, and PD severity over an average of 9 years of follow-up, utilizing a machine learning algorithm to quantify disease severity based on annual antiparkinsonian medication purchases. Results: Our analysis revealed a significant reduction in PD occurrence following tetanus–diphtheria (Td) vaccination, with an adjusted odds ratio of 0.17 (95% CI [0.04, 0.70]) for PD onset within 5 years post-vaccination. Furthermore, a time-dependent relationship was identified between the duration since vaccination and both the rate of PD onset and disease progression. Notably, we observed that antimicrobial treatments significantly influenced disease severity, consistent with the antibiotic sensitivity profile of Clostridium tetani. Conclusions: These findings support the hypothesis that tetanus vaccination and/or C. tetani eradication may reduce PD occurrence and slow its progression, suggesting promising directions for future research in PD prevention and treatment. [ABSTRACT FROM AUTHOR]

Horses are exquisitely sensitive to tetanus neurotoxin and are exposed to the risk of infection with Clostridium tetani throughout life. The vaccine against tetanus is highly effective at preventing disease, whereas tetanus in unvaccinated populations is associated with high mortality rates. Current guidelines in New Zealand and Australia for the available vaccine contain contradictions and limitations surrounding the optimal tetanus immunisation protocols for both adult horses and foals. This review critically evaluates the scientific literature on tetanus prophylaxis in horses within the context of equine practice and available products in New Zealand and Australia. The review was conducted by a panel of industry and specialist veterinarians to obtain agreement on nine equine tetanus prophylaxis guidelines for practising veterinarians. The primary protocol for tetanus toxoid (TT) immunisation consists of a three-dose series IM for all horses ≥ 6 months of age, and a four-dose series IM is proposed if commencing vaccination in foals between 3 and 6 months of age. Tetanus prophylaxis in foals < 3 months of age relies on passive immunity strategies. Following the completion of the primary protocol, a TT booster dose IM should be administered within 5 years, and every 5 years thereafter. When followed, these protocols should provide adequate protection against tetanus in horses. Additional tetanus prophylaxis guidelines are provided for veterinarians attending a horse experiencing a known "risk event" (e.g. wound, hoof abscess, surgery, umbilical infection). When a correctly vaccinated horse experiences a risk event, pre-existing immunity provides protection against tetanus. When an unvaccinated horse or one with unknown vaccination status, or a foal born to an unvaccinated dam, experiences a risk event, TT IM and tetanus antitoxin (TAT) 1,500 IU SC should be administered simultaneously at separate sites, and the TT primary immunisation protocol should subsequently be completed for the horse's respective age. In previously immunised pregnant broodmares, a TT booster dose administered 4–8 weeks prior to parturition optimises the transfer of passive immunity against tetanus to the newborn foal via colostrum; provided that post-natal IgG concentration in serum is > 800 mg/dL (8 g/L), such foals should be passively protected against tetanus up to 6 months of age. Survivors of clinical tetanus must still receive the primary protocol for vaccination against tetanus. In summary, all horses in New Zealand and Australia should be vaccinated against tetanus with protection maintained throughout life via TT booster doses, facilitated by accurate medical record keeping and client education. [ABSTRACT FROM AUTHOR]

Abstract Tetanus has become an increasingly rare infectious disease due to the development of successful vaccination programs in the mid-20th century. In resource-rich countries, mainly unvaccinated or partly vaccinated risk groups are affected, whereas tetanus still remains prevalent in resource-limited countries. The decreasing incidence in developed countries has hindered clinical trials evaluating the best treatment modalities for tetanus infections. Current guidelines are based on a small number of studies and case reports. So far, these studies have shown potential benefits of treating tetanus infections with benzodiazepines, magnesium sulfate and baclofen. Additionally, several treatments have been shown to be useful in stabilizing and supporting patients with tetanus. However, each treatment modality has limitations, from negative side effects to logistical challenges, especially in developing countries. Therefore, further knowledge is required to evaluate the best use of each treatment and to further optimize patient care. This knowledge can contribute to the reduction of the burden of disease in countries where tetanus remains prevalent and where resources are limited, though vaccination is the most effective method to achieve this. This case report describes the treatment of a Dutch patient with tetanus infection and illustrates the role of benzodiazepines as well as other key aspects of treating patients with tetanus.

Tetanus is a toxigenic illness caused by the action of Clostridium tetani neurotoxin (TeNT), which results in partial or generalized muscle stiffness in infected mammals and birds. The disease is rarely reported in cats due to their innate resistance to the toxin. This multicentric retrospective study aimed to describe a significant population of cats with a diagnosis of tetanus and report their signalment, clinical and neurological signs, diagnostic findings, treatment, and outcome. A retrospective search through medical records from 11 referral centers in Europe resulted in the identification of 27 cases of feline tetanus from July 2005 to April 2023. These cases were further compared with previously reported cases in the veterinary literature. Young cats were more commonly represented than older cats, with a median age of 4 years. Clinical signs were initially characterized by a lame and/or stiff limb, near the primary injury site, in 17/26 (65%) cats. Signs were focal or multifocal in 21/27 (78%) cats of this study and one typical sign was the inability to flex the most severely affected limbs. Electrodiagnostic studies revealed characteristic changes, such as continuous spontaneous motor unit discharges in both agonist and antagonist muscles. Such studies are particularly useful in focal and multifocal cases and should be performed to further support the diagnosis. The toxin was successfully identified in one case using the mouse bioassay. Treatment included antibiotherapy (metronidazole) in most cases, muscle relaxants, appropriate nursing cares and handling of potential complications. Overall, the outcome appeared to be positive, with only 1/27 (3.7%) cats being euthanized due to financial restrains. 23/25 (92%) cats returned to an independent ambulatory capacity on all limbs within a median delay of 25 days. Mild to moderate longterm sequelae were reported in eight (30%) cats. This multicentric study is the first to bring together such a large number of cats affected with tetanus. Presentation of the disease in cats differs from that observed in humans and dogs, with most cats being locally affected. Compared to previous reports of tetanus, this series of cats had a better outcome overall, especially for cats affected with generalized tetanus. [ABSTRACT FROM AUTHOR]

Traditionally, diphtheria-tetanus-pertussis (DTwP or DTaP) as pediatric vaccines are produced from the corresponding inactivated toxins or whole -cell pathogenic bacteria of Corynebacterium diphtheria toxin (Tox), Clostridium tetani toxin (TetX) and Bordetella pertussis. There are major concerns in the classic or acellular DTP (DTaP) vaccine production processes from native live bacterial sources as it may raise concerns on adverse effects and safety issues, complexity of the purifications for each agent as well as cost. Here, we designated a recombinant multi-epitope vaccine candidates by vaccinoinformatics study to address the mentioned issues and to develop a single trivalent fusion protein as a potent recombinant DTP vaccine. To achieve these goals, stages of immunebioinformatics were retrieved using proteinaceous toxins sequences, predicting secondary/tertiary structure and transmembrane topology, energy minimization, and model validation. Then, conformational and linear Bcell epitope prediction by several servers, mapping of consensus linear/discontinuous immunogenic regions and construction synthetic fusion vaccine candidates in respect to optimal immunogenic, physicochemical properties and high expression in prokaryote host were achieved. Finally, reverse translation, codon optimization, addition of cloning tags for pet 28a vector and optimization of physicochemical properties of synthetic trivalent fusion protein were performed. Through various hybrid immuno-informatics and structural bioinformatics analysis of predicted and experimental epitopes finally, 12 new consensus highly immunogenic linear and discontinuous epitopes in Tox, TetX and PTXa proteins were selected. The peptide sequences of these immunogenic regions were as follows: PTXA (AA34-64, AA184-256 and AA98-116), Tox (AA47-76, AA117-159, AA515- 557 and AA245-265) and TetX (AA226-249, AA819-844, AA923-967, AA1009-1067 and AA1225-1315). In addition, the characteristics of the recombinant trivalent fusion construct were; 546 residue length, soluble (Grand average of hydropathicity (GRAVY) was -0.475), estimated half-life was >10 hours in Escherichia coli, pI 5.94 (a little acidic), stable protein (The instability index (II) 35.58) as well as thermally stable (Aliphatic index (AI) 71.67). The putative antigenic epitopes from different organisms in a single protein, as in the current study, will possibly improve the protective efficacy as novel potent, safe, cheap and broad-spectrum vaccines for better prevention of diphtheria, tetanus and pertussis infections in the future. [ABSTRACT FROM AUTHOR]

Tetanus has become an increasingly rare infectious disease due to the development of successful vaccination programs in the mid-20th century. In resource-rich countries, mainly unvaccinated or partly vaccinated risk groups are affected, whereas tetanus still remains prevalent in resource-limited countries. The decreasing incidence in developed countries has hindered clinical trials evaluating the best treatment modalities for tetanus infections. Current guidelines are based on a small number of studies and case reports. So far, these studies have shown potential benefits of treating tetanus infections with benzodiazepines, magnesium sulfate and baclofen. Additionally, several treatments have been shown to be useful in stabilizing and supporting patients with tetanus. However, each treatment modality has limitations, from negative side effects to logistical challenges, especially in developing countries. Therefore, further knowledge is required to evaluate the best use of each treatment and to further optimize patient care. This knowledge can contribute to the reduction of the burden of disease in countries where tetanus remains prevalent and where resources are limited, though vaccination is the most effective method to achieve this. This case report describes the treatment of a Dutch patient with tetanus infection and illustrates the role of benzodiazepines as well as other key aspects of treating patients with tetanus. [ABSTRACT FROM AUTHOR]

Case summaryA 3-month-old Airedale dog with clinically diagnosed generalized tetanus was investigated for the occurrence of excessive paddling and chewing movements when sleeping. Electroencephalogram (EEG) with time-locked video over 31 hours determined occurrence of the abnormal movements to be within 20 to 180 seconds of the onset of rapid eye movement (REM) sleep, but not at any other stage of wakefulness or sleep. No epileptiform activity was noted. Clinical signs of generalized tetanus resolved over 8 weeks with antimicrobial and symptomatic treatment, and sleep-associated movements resolved 6 weeks after presentation.Clinical relevanceRapid eye movement sleep behavior disorder (RBD) has been suspected in dogs with generalized tetanus but not confirmed by correlation of repeated episodes of vocalization or motor behaviors or both with REM sleep defined by an EEG. The case further defines RBD in dogs with tetanus, and highlights the value of EEG to differentiate among different parasomnias and epileptiform activity.

Tetanus neurotoxins (TeNT) and botulinum neurotoxins (BoNTs) are closely related ~150 kDa protein toxins that together comprise the group of clostridial neurotoxins (CNTs) expressed by various species of Clostridia. While TeNT is expressed as a single polypeptide, BoNTs are always produced alongside multiple non-toxic proteins that form a stabilizing complex with BoNT and are encoded in a conserved toxin gene cluster. It is unknown how tent evolved without a similar gene cluster and why complex-free TeNT is secreted as a stable and soluble protein by C. tetani, whereas complexing proteins appear to be essential for BoNT stability in culture supernatants of C. botulinum. To assess whether the stability of TeNT is due to an innate property of the toxin or is a result of C. tetani’s intra- and extra-cellular environment, both TeNT and complex-free BoNT/A1ERY were expressed recombinantly in atoxic C. tetani and analyzed for expression and stability. The strong clostridial ferredoxin (fdx) promotor resulted in the expression of recombinant TeNT at greater levels and earlier time points than endogenously produced TeNT. Recombinant BoNT/A1ERY was similarly expressed by atoxic C. tetani, although partial degradation was observed. The rBoNT/A1ERY produced in C. tetani was also partially proteolytically processed to the dichain form. Investigations of bacterial growth media and pH conditions found that the stability of rTeNT and rBoNT/A1ERY in spent media of C. tetani or C. botulinum was affected by growth media but not by pH. These data indicate that the distinct metabolism of C. tetani or C. botulinum under various growth conditions is a primary factor in creating a more or less favorable environment for complex-free CNT stability.

Tetanus is a toxigenic illness caused by the action of Clostridium tetani neurotoxin (TeNT), which results in partial or generalized muscle stiffness in infected mammals and birds. The disease is rarely reported in cats due to their innate resistance to the toxin. This multicentric retrospective study aimed to describe a significant population of cats with a diagnosis of tetanus and report their signalment, clinical and neurological signs, diagnostic findings, treatment, and outcome. A retrospective search through medical records from 11 referral centers in Europe resulted in the identification of 27 cases of feline tetanus from July 2005 to April 2023. These cases were further compared with previously reported cases in the veterinary literature. Young cats were more commonly represented than older cats, with a median age of 4 years. Clinical signs were initially characterized by a lame and/or stiff limb, near the primary injury site, in 17/26 (65%) cats. Signs were focal or multifocal in 21/27 (78%) cats of this study and one typical sign was the inability to flex the most severely affected limbs. Electrodiagnostic studies revealed characteristic changes, such as continuous spontaneous motor unit discharges in both agonist and antagonist muscles. Such studies are particularly useful in focal and multifocal cases and should be performed to further support the diagnosis. The toxin was successfully identified in one case using the mouse bioassay. Treatment included antibiotherapy (metronidazole) in most cases, muscle relaxants, appropriate nursing cares and handling of potential complications. Overall, the outcome appeared to be positive, with only 1/27 (3.7%) cats being euthanized due to financial restrains. 23/25 (92%) cats returned to an independent ambulatory capacity on all limbs within a median delay of 25 days. Mild to moderate long-term sequelae were reported in eight (30%) cats. This multicentric study is the first to bring together such a large number of cats affected with tetanus. Presentation of the disease in cats differs from that observed in humans and dogs, with most cats being locally affected. Compared to previous reports of tetanus, this series of cats had a better outcome overall, especially for cats affected with generalized tetanus.

Jun Hirai,1,2 Nobuaki Mori,1,2 Yuichi Shibata,2 Nobuhiro Asai,1,2 Mao Hagihara,3 Hiroshige Mikamo1,2 1Department of Clinical Infectious Diseases, Aichi Medical University Hospital, Nagakute, Aichi, Japan; 2Department of Infection, Prevention, and Control, Aichi Medical University Hospital, Nagakute, Aichi, Japan; 3Department of Molecular Epidemiology and Biomedical Sciences, Aichi Medical University, Nagakute, Aichi, JapanCorrespondence: Jun Hirai, Department of Clinical Infectious Diseases, Aichi Medical University Hospital, 1-1, Yazako-karimata, Nagakute, Aichi, 480-1195, Japan, Tel +81-561-62-3311, Fax +81-561-76-2673, Email hiraichimed@gmail.comBackground: The incidence of tetanus has significantly declined in developed countries owing to widespread vaccination efforts. However, it remains a threat worldwide, including in Japan, because of the sharp decline in antibody titers against tetanus in adults. Animal bites, including canine bites, are potential sources of tetanus infection. This case highlights the rarity of tetanus caused by canine bites and the need for continued vigilance for tetanus prevention. This case report and literature review aimed to shed light on the clinical course and outcomes of tetanus following a canine bite.Case Presentation: A 46-year-old Japanese man with no medical history presented with symptoms of tetanus, such as difficulty in opening his mouth, 19 days after a canine bite on his right hand. He was born and brought up in Japan. He had never been vaccinated against tetanus. Despite washing the wound and receiving human tetanus immunoglobulin (HTIG) and a tetanus toxoid vaccine, the patient developed tetanus. After intravenous metronidazole and HTIG were administered, the symptoms improved gradually. The patient was discharged after a 12-day hospital stay.Discussion: This is the first reported case of canine bite-induced tetanus in Japan, where tetanus toxoid vaccination is provided routinely. This case highlights the waning immunity in adults and the critical need for education on tetanus immunization, including catch-up immunization, particularly for adults and individuals in high-risk occupations. A review of the existing literature revealed only four cases of tetanus following canine bites between 1889 and 2018. All patients experienced symptom onset between 3 and 19 days post injury. Treatment typically involved HTIG, metronidazole, and toxoid administration. A higher risk of mortality is seen in unvaccinated individuals than in vaccinated individuals, highlighting the critical role of tetanus vaccination.Conclusion: Physicians should consider canine bite-induced tetanus in the differential diagnosis when patients exhibit relevant symptoms.Keywords: tetanus, Clostridium tetani, canine, bite, vaccine, Japan

Background: Parkinson’s disease (PD) is a neurodegenerative disorder that progressively damages the autonomic and central nervous systems, leading to hallmark symptoms such as resting tremor, bradykinesia, and rigidity. Despite extensive research, the underlying cause of PD remains unclear, and current treatments are unable to halt the progression of the disease. In this retrospective study, based on historical electronic health records (EHR) from a national health provider covering the period from 2003 to 2023, we investigated the impact of vaccination and medication purchases on PD occurrence and severity. Methods: Using a case–control design, we compared the vaccination histories of 1446 PD patients with 7230 matched controls to assess the association between vaccination and PD onset. Additionally, we explored statistical associations between vaccination, medication purchases, and PD severity over an average of 9 years of follow-up, utilizing a machine learning algorithm to quantify disease severity based on annual antiparkinsonian medication purchases. Results: Our analysis revealed a significant reduction in PD occurrence following tetanus–diphtheria (Td) vaccination, with an adjusted odds ratio of 0.17 (95% CI [0.04, 0.70]) for PD onset within 5 years post-vaccination. Furthermore, a time-dependent relationship was identified between the duration since vaccination and both the rate of PD onset and disease progression. Notably, we observed that antimicrobial treatments significantly influenced disease severity, consistent with the antibiotic sensitivity profile of Clostridium tetani. Conclusions: These findings support the hypothesis that tetanus vaccination and/or C. tetani eradication may reduce PD occurrence and slow its progression, suggesting promising directions for future research in PD prevention and treatment.

A 10-month-old, 3  kg, female spayed Domestic Shorthair cat was presented with a chronic, infected wound at the level of the proximo-lateral left pelvic limb. General physical examination revealed a weight-bearing lameness of the left pelvic limb, which was moderately and circumferentially swollen and edematous proximal to the tarsal joint. On the lateral aspect of the proximal thigh, there was a chronic wound of 1  cm in diameter and an additional exudative skin lesion was present throughout the whole length of the caudo-lateral thigh. Complete blood count and serum biochemistry profile revealed mild anemia, increased serum amyloid A, hyponatraemia, hypochloraemia, hypocalcaemia, hyperkalaemia, hypermagnesaemia, hyperglycaemia, increased creatine kinase, and increased liver parameters. Surgical exploration of the wound was performed, and necrotizing fasciitis was suspected. The affected limb was amputated and swabs for bacterial culture were taken from both the skin lesions and surgical site before wound closure. One day after surgery, mild muscular contractions on the forehead and an increased muscle tone of the right pelvic limb were evident. One day later, the cat developed a generalized increase in extensor tone, with intermittent opisthotonos, resulting in lateral recumbency. Based on these clinical signs, a diagnosis of generalized tetanus was made and treatment with midazolam, methocarbamol, and metronidazole was started. Despite an improvement of all blood parameters, the cat progressively deteriorated and 4  days after surgery, it developed episodes of tetanic convulsions, associated with hyperthermia and ventricular arrhythmias. Despite intensive care and medical management, the cat died following a cardio-respiratory arrest 3  days later. This case report describes a rare case of severe generalized tetanus in a young cat. [ABSTRACT FROM AUTHOR]

Objective To evaluate the seasonality of canine tetanus in England.Methods Medical records of a single referral hospital in England were reviewed. Dogs diagnosed with localized or generalized tetanus between January 2006 and June 2017 were studied.Results Forty-nine cases were included. The prevalence of tetanus in England was significantly higher in the winter when compared with the summer (P = 0.002) and autumn (P = 0.024), with the highest number of cases recorded in February.Conclusions The prevalence of canine tetanus in England was significantly higher in winter months, especially in February.

A 10-month-old, 3 kg, female spayed Domestic Shorthair cat was presented with a chronic, infected wound at the level of the proximo-lateral left pelvic limb. General physical examination revealed a weight-bearing lameness of the left pelvic limb, which was moderately and circumferentially swollen and edematous proximal to the tarsal joint. On the lateral aspect of the proximal thigh, there was a chronic wound of 1 cm in diameter and an additional exudative skin lesion was present throughout the whole length of the caudo-lateral thigh. Complete blood count and serum biochemistry profile revealed mild anemia, increased serum amyloid A, hyponatraemia, hypochloraemia, hypocalcaemia, hyperkalaemia, hypermagnesaemia, hyperglycaemia, increased creatine kinase, and increased liver parameters. Surgical exploration of the wound was performed, and necrotizing fasciitis was suspected. The affected limb was amputated and swabs for bacterial culture were taken from both the skin lesions and surgical site before wound closure. One day after surgery, mild muscular contractions on the forehead and an increased muscle tone of the right pelvic limb were evident. One day later, the cat developed a generalized increase in extensor tone, with intermittent opisthotonos, resulting in lateral recumbency. Based on these clinical signs, a diagnosis of generalized tetanus was made and treatment with midazolam, methocarbamol, and metronidazole was started. Despite an improvement of all blood parameters, the cat progressively deteriorated and 4 days after surgery, it developed episodes of tetanic convulsions, associated with hyperthermia and ventricular arrhythmias. Despite intensive care and medical management, the cat died following a cardio-respiratory arrest 3 days later. This case report describes a rare case of severe generalized tetanus in a young cat.

ABSTRACTClostridium tetani is the etiological agent of the disease tetanus, resulting in various symptoms, including neck tension, tongue-twisting, a decline in walking ability, and death in severe cases. To date, publicly available genome sequences of C. tetani are limited. C. tetani was isolated from soil in Kumamoto Prefecture, Japan, between 2020 and 2022. Subsequently, bacteriological, biochemical, genetic, and immunological tests were conducted. Among the 46 soil sampling sites, 151 C. tetani strains were isolated from 33 sites, with a 71.7% positivity rate. Comparative genomic analysis subdivided clade 1 into clades 1-1, 1-2, and 1-3. The enzyme-linked immunosorbent assay revealed that partial clade 1-3 strains produced over sevenfold more tetanus neurotoxin (TeNT) than the reference strain (first isolate of C. tetani KHSU-154301-001). RNA-Seq analysis supported this elevated tetanus toxin tetX gene expression. Additionally, tetX-negative strains were isolated from the clade, indicating that TeNT may not be essential for ensuring the survival of their offspring without its representative toxin-causing tetanus. Five strains acquired the tetracycline resistance gene tet(M) in the tetX plasmid, resulting in significantly reduced susceptibility to tetracycline. In conclusion, although we investigated C. tetani isolates from a specific region of the Kumamoto Prefecture, we demonstrated that these isolates exhibit variable genomic features, toxin productivity, and substantial acquired antimicrobial resistance. These findings indicate that multiple C. tetani lineages have successfully adapted to the soil in Japan, as evidenced by the additional genome information and toxin productivity assays in this study.IMPORTANCEC. tetani is a spore-forming, anaerobic bacterium that produces a toxin causing muscle stiffness and paralysis. Tetanus is preventable with the toxoid vaccine, but it remains a significant public health threat in regions with low vaccine coverage. However, there are relatively few isolates and limited genomic information available worldwide. In Japan, about 100 cases are reported each year, but there have been no nationwide surveys of isolates, and no genomic information from Japanese isolates has been published. In our study, we analyzed the genomes of 151 strains from a limited survey of soil in Kumamoto, Japan. Our findings revealed a high degree of genetic diversity, and we also identified a subset of strains that produced significantly more toxin, which provides new insights into the pathogenesis of tetanus. Our findings lay the foundation for future studies to investigate the distribution and evolution of C. tetani in Japan and neighboring countries.

The worsening of antibiotic resistance is a multifactorial process. One aspect of this is the counterfeiting of antibiotic medications. This is supposed to be particularly high in developing countries, including Nigeria. Therefore, the potency of some antibiotic drugs dispensed in community pharmacies in Gwale, Kano, Nigeria, was investigated in this case study. Three products, each from different manufacturers, with the active ingredients of ceftriaxone, gentamicin, ciprofloxacin, and metronidazole, respectively, were included in this study. By means of a disc-diffusion assay, the effect against the typed strains Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) as well as Clostridium tetani isolated from soil was tested. Clinical isolates of S. aureus and E. coli were also used. While antibiotics, with the exception of ciprofloxacin-containing preparations against C. tetani, showed acceptable efficacy against the typed strains by comparison with the clinical science laboratory references, a predominant failure was observed with the clinical isolates. Thus, the investigated drug preparations can be considered of acceptable quality for the treatment of susceptible bacterial infections. This excludes counterfeits in the sampled preparations. However, the insufficient efficacy against clinical isolates further documents the severity of nosocomial bacteria. [ABSTRACT FROM AUTHOR]

Tetanus is an infectious disease caused by Clostridium tetani toxin. Although easily preventable through vaccination, over 73,000 new infections and 35,000 deaths due to tetanus occurred worldwide in 2019, with higher rates in countries with healthcare barriers. Here, we present a clinical case of C. tetani infection in an 85-year-old patient. Patient robustness and high functional reserve before infection are favorable predictors of survival for an otherwise fatal disease. However, the patient did not experience any severe complications. Therefore, this report is a strong call for tetanus vaccination. [ABSTRACT FROM AUTHOR]

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Abstract Background Tetanus remains a severe life-threatening infectious disease and neurological disorder in many parts of the world, where immunization programs are suboptimal. Any human injury or trauma has the possibility of getting infected with Clostridium tetani which is the sole causative bacterium of tetanus. Evidence is available that TAT may cause anaphylaxis and late serum sickness, while there has been no study conducted in Ethiopia. The Ethiopian Ministry of Health standard treatment guideline recommends tetanus prophylaxis for all tetanus-prone wounds. This study aimed to evaluate the safety of TAT administration in adults exposed to tetanus-prone wounds in Ethiopia. Methods The target product of this study was the equine tetanus antitoxin developed and manufactured by the ViNS Bioproducts Limited, India (Code: 130202084, A.W.No: 15/AAW/PI/02.00, DT: 25.04.2016). The product is delivered with the dose of 1000/1500 IU intramuscularly or subcutaneously to individuals at risk of tetanus infection for prophylactic purposes. The study was carried out in 11 healthcare facilities in Addis Ababa, Ethiopia, that had a relatively high clients load for tetanus-prone wounds. Medical records of patients with tetanus-prone wounds who received the equine TAT were reviewed retrospectively for any adverse events following immunization according to the World Health Organization (WHO) definition for adverse events following immunization (AEFI). Results There were more than 20,000 patients treated for trauma in the facilities from 2015 to 2019. Upon revision of available registration books, we identified 6000 charts to be eligible for the study, of which 1213 charts that had complete and reliable data on the AEFI profile of the TAT were included in the final analysis. The median age of the study participants was 26 years (IQR = 11 years, age range: 18–91 years) and 78% (949) were male. The tetanus-prone wounds resulted mainly from stab (44%, 535) and blunt force (30%, 362), and the most common sites of wounds were hand (22%, 270) and head (21%, 253). The most and least frequently occurring types of wounds were open wounds (77%, 930) and organ system injury (0.003%, 4), respectively. The mean time of presenting at health facilities from the onset of trauma was 2.96 h. Of the total 1231 participants, one male participant who presented within 3 h after experiencing a wound on his nose at the workplace had a severe local reaction immediately after injection of the TAT. No AEFI was recorded for the other participants. Conclusions The adverse event following immunization of the equine tetanus antitoxin produced by the ViNS Bioproducts Limited was very rare. A regular review of the product’s safety performance and systematic collection and analysis of adverse event reports are important to ensure the safety of the product.

Abstract Case Summary A 3‐month‐old Airedale dog with clinically diagnosed generalized tetanus was investigated for the occurrence of excessive paddling and chewing movements when sleeping. Electroencephalogram (EEG) with time‐locked video over 31 hours determined occurrence of the abnormal movements to be within 20 to 180 seconds of the onset of rapid eye movement (REM) sleep, but not at any other stage of wakefulness or sleep. No epileptiform activity was noted. Clinical signs of generalized tetanus resolved over 8 weeks with antimicrobial and symptomatic treatment, and sleep‐associated movements resolved 6 weeks after presentation. Clinical Relevance Rapid eye movement sleep behavior disorder (RBD) has been suspected in dogs with generalized tetanus but not confirmed by correlation of repeated episodes of vocalization or motor behaviors or both with REM sleep defined by an EEG. The case further defines RBD in dogs with tetanus, and highlights the value of EEG to differentiate among different parasomnias and epileptiform activity.

Highlight: • Tetanus in patients with infected sutured wound. • Complications of death in generalized tetanus. • The importance of education about tetanus. ABSTRACT Introduction: Tetanus is a disease caused by Clostridium tetani, which produces an exotoxin. Tetanus can rapidly progress into life-threatening muscle spasms accompanied by respiratory insufficiency with or without autonomic dysfunction. Case: A 43-year-old male patient was referred from the public health center to Tora Belo Hospital with mouth stiffness and difficulty swallowing food. He also had stiffness and pain in the neck, abdomen, and lower back. Seven days prior, the patient received lacerations on his right forearm caused by a machete. The patient’s wound had been stitched up without an anti-tetanus injection at the public health center. The symptoms worsened on the second day of admission, and the patient died from respiratory failure. Conclusion: It is very important to enhance public awareness that tetanus is a preventable disease, with proper vaccination and sanitization potentially lowering mortality and morbidity.

ObjectiveTetanus is a severe neurologic disease caused by Clostridium tetani, resulting in spastic paralysis. Canine tetanus is associated with serious complications such as aspiration and a high mortality rate of up to 50%.Materials and methodsMedical records of all dogs diagnosed with tetanus over 8 years (2014–2022) were analyzed for severity grade, treatment protocols, nutritional management, and complications, as well as outcome, vaccination, and antibody production in some dogs. No medical records were excluded. Normality was analyzed by the D'Agostino–Pearson test. Parametric, normally distributed data were presented as mean ± standard deviation. Non-parametric, non-normally distributed data were presented as median (m) and range (minimum–maximum). The association between tetanus grade, progression of diseases, and duration of hospitalization was analyzed using the t-test, Mann–Whitney U test, and Kruskal–Wallis test. A P ≤ 0.05 was considered significant.ResultsEighteen dogs were identified. Most affected dogs were classified into severity grade II (66.7%, 12 of 18). Clinical signs deteriorated in 55.6% of dogs (10 of 18). A source was identified in 88.9% of dogs (16 of 18). Nine dogs required surgical wound revision. A percutaneous endoscopic gastropexy tube was placed in 83.3% of dogs (15 of 18) for nutritional support. Medical treatment included metronidazole, methocarbamol, and combinations of different sedatives adapted to the patient's requirements. Tetanus antitoxin was used in 72.2% of dogs (13 of 18) without reported adverse events. The survival rate was 88.9% (16 of 18). Complications, such as hypertension, aspiration pneumonia, and laryngeal spasm occurred in 12 of 18 dogs. Median hospitalization time (8 days; range 0–16 days) was associated with the maximum tetanus severity grade (p = 0.022). Rapid eye movement behavior disorder was observed in 72.2% of dogs (13 of 18). In 5 dogs, antibodies were measured after recovery, and in 4 of 5 dogs, no antibodies were detectable despite generalized tetanus disease. Vaccination with tetanus toxoid was performed in five dogs following the disease.ConclusionIn the present study, the mortality rate was lower than previously reported. Tetanus is still a life-threatening disease, but the prognosis may be good if adequate management and monitoring can be ensured.

Tetanus is an acute infectious disease, caused by spores of the bacterium Clostridium tetani. The spores are found everywhere in the environment, particularly in soil, ash, intestinal tracts/faeces of animals and humans, on the surfaces of skin and rusty tools like nails, needles, barbed wires, etc. Being very resistant to heat and most antiseptics, the spores can survive for years. Anyone can get tetanus, but the disease is particularly common and serious in new born babies and pregnant women, who have not been sufficiently immunised with Tetanus Toxoid (TT) containing vaccines. The disease remains an important public health problem in many parts of the world, but especially in low income countries or districts, where immunisation coverage is low and unclean birth practices are common. There are no particular diagnostic tests for tetanus, the diagnosis is made clinically. Hence, early identification of cases of tetanus becomes important for early intervention. Herein, the authors reported four cases (1 male and 3 females) of tetanus, with varied clinical manifestations, management and outcome. The patients were diagnosed with severe generalised tetanus, otogenic tetanus, mild generalised tetanus and severe generalised tetanus respectively. The patients were treated with inj. diazepam, inj. tetanus toxoid, inj. methocarbamol. Two of them recovered completely and two could not be revived.

Tetanus is an acute and often fatal infectious disease caused by Clostridium tetani. Tetanus toxin (TT) is responsible for spastic paralysis observed in tetanus. Anti-tetanus antibodies obtained from horses and humans are the most antitoxins used for tetanus treatment, although some clinical side effects and disadvantages have been reported in their application. The aim of this study is the production of anti-TT IgY and evaluation of its protective effects in a mouse model. Anti-TT IgY was purified from the egg yolk using PEG6000 precipitation and water dilution methods, and its purity was verified by SDS-PAGE. Finally, the potency of purified anti-TT IgY in neutralizing the lethal effects of TT was studied in vivo using a mouse model. PEG6000 precipitation method had better results. Animal studies showed that the purified IgY neutralized the toxic effects of 100 MLD of TT and multiple intravenous-dose injections of anti-TT IgY also had a continuous effect of TT neutralization. The purified anti-TT IgY was effective in neutralizing the lethal activity of TT in a mouse model. Our results suggested that IgY could be an alternative therapeutic source for the management of tetanus in the future. Abbreviations Anti-TT, Anti-tetanus toxin; ELISA, Enzyme-linked immunosorbent assay; IgY, Immunoglobulin Y; MLD, Minimum lethal dose; PBS, Phosphate buffer solution; PEG, Polyethylene glycol; SDS-PAGE, Sodium dodecyl sulfate polyacrylamide gel electrophoresis; TIG, Tetanus immune globulin; TT, Tetanus toxin; WD, Water dilution; RT, Room temperature. [ABSTRACT FROM AUTHOR]

O tétano é uma doença infecciosa não contagiosa, causada por exotoxinas produzidas por Clostridium tetani (C. tetani), um microrganismo de distribuição mundial, gram-positivo, encontrado sob a forma vegetativa ou esporulada. O objetivo do presente trabalho é apresentar o relato de caso de um equino, macho, adulto, com 450 kg de peso corporal, sem histórico de vacinação e acometido por tétano. O animal foi atendido no Hospital Universitário de Medicina Veterinária da Universidade Federal do Recôncavo da Bahia, e apresentou ao exame clínico trismo mandibular, andar rígido, musculatura contraída e prolapso de terceira pálpebra. A partir do diagnóstico clínico, instituiu-se o tratamento com soro antitetânico, penicilina, acepromazina 1%, fluidoterapia e suplementação com soluções energéticas e vitaminas. Foram realizados ainda, o debridamento da lesão do casco e enema para a remoção das fezes da ampola retal. No décimo primeiro dia de internamento o animal apresentou significativa melhora clínica, e após 29 dias de internamento o animal teve alta médica. O diagnóstico precoce associado ao manejo do local de infecção e o tratamento intensivo foram fatores que contribuíram para a recuperação deste animal.

Tetanus is extremely rare in developed countries. We report the first documented case of tetanus in the province of Alberta since 2016: a farmer that developed trismus, shoulder stiffness, and fevers eight days following orthopedic surgery. Tetanus immunoglobulin elicited rapid recovery. We highlight risk factors, pathogenesis, epidemiology, and diagnostic challenges.

The worsening of antibiotic resistance is a multifactorial process. One aspect of this is the counterfeiting of antibiotic medications. This is supposed to be particularly high in developing countries, including Nigeria. Therefore, the potency of some antibiotic drugs dispensed in community pharmacies in Gwale, Kano, Nigeria, was investigated in this case study. Three products, each from different manufacturers, with the active ingredients of ceftriaxone, gentamicin, ciprofloxacin, and metronidazole, respectively, were included in this study. By means of a disc-diffusion assay, the effect against the typed strains Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) as well as Clostridium tetani isolated from soil was tested. Clinical isolates of S. aureus and E. coli were also used. While antibiotics, with the exception of ciprofloxacin-containing preparations against C. tetani, showed acceptable efficacy against the typed strains by comparison with the clinical science laboratory references, a predominant failure was observed with the clinical isolates. Thus, the investigated drug preparations can be considered of acceptable quality for the treatment of susceptible bacterial infections. This excludes counterfeits in the sampled preparations. However, the insufficient efficacy against clinical isolates further documents the severity of nosocomial bacteria.

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Clostridium is a genus comprising Gram-positive, rod-shaped, spore-forming, anaerobic bacteria that cause a variety of diseases. However, there is a shortage of information regarding antibiotic resistance in the genus in Saudi Arabia. This comprehensive analysis of research results published up until December 2021 intends to highlight the incidence of antibiotic resistance in Clostridium species in Saudi Arabia. PubMed, Google Scholar, Web of Science, SDL, and ScienceDirect databases were searched using specific keywords, and ten publications on antibiotic resistance in Clostridium species in Saudi Arabia were identified. We found that the rates of resistance of Clostridium difficile to antibiotics were as follows: 42% for ciprofloxacin, 83% for gentamicin, 28% for clindamycin, 25% for penicillin, 100% for levofloxacin, 24% for tetracycline, 77% for nalidixic acid, 50% for erythromycin, 72% for ampicillin, and 28% for moxifloxacin; whereas those of C. perfringens were: 21% for metronidazole, 83% for ceftiofur, 39% for clindamycin, 59% for penicillin, 62% for erythromycin, 47% for oxytetracycline, and 47% for lincomycin. The current findings suggest that ceftiofur, erythromycin, lincomycin, and oxytetracycline should not be used in C. perfringens infection treatments in humans or animals in Saudi Arabia. [ABSTRACT FROM AUTHOR]

Tetanus is an acute infectious disease, caused by spores of the bacterium Clostridium tetani. The spores are found everywhere in the environment, particularly in soil, ash, intestinal tracts/faeces of animals and humans, on the surfaces of skin and rusty tools like nails, needles, barbed wires, etc. Being very resistant to heat and most antiseptics, the spores can survive for years. Anyone can get tetanus, but the disease is particularly common and serious in new born babies and pregnant women, who have not been sufficiently immunised with Tetanus Toxoid (TT) containing vaccines. The disease remains an important public health problem in many parts of the world, but especially in low income countries or districts, where immunisation coverage is low and unclean birth practices are common. There are no particular diagnostic tests for tetanus, the diagnosis is made clinically. Hence, early identification of cases of tetanus becomes important for early intervention. Herein, the authors reported four cases (1 male and 3 females) of tetanus, with varied clinical manifestations, management and outcome. The patients were diagnosed with severe generalised tetanus, otogenic tetanus, mild generalised tetanus and severe generalised tetanus respectively. The patients were treated with inj. diazepam, inj. tetanus toxoid, inj. methocarbamol. Two of them recovered completely and two could not be revived. [ABSTRACT FROM AUTHOR]

Abstract Background Tetanus remains common in many low- and middle-income countries, but as critical care services improve, mortality from tetanus is improving. Nevertheless, patients develop severe syndromes associated with autonomic nervous system disturbance (ANSD) and the requirement for mechanical ventilation (MV). Understanding factors associated with worse outcome in such settings is important to direct interventions. In this study, we investigate risk factors for disease severity and long-term physical outcome in adults with tetanus admitted to a Vietnamese intensive care unit. Methods Clinical and demographic variables were collected prospectively from 180 adults with tetanus. Physical function component scores (PCS), calculated from Short Form Health Survey (SF-36), were assessed in 79 patients at hospital discharge, 3 and 6 months post discharge. Results Age, temperature, heart rate, lower peripheral oxygen saturation (SpO2) and shorter time from first symptom to admission were associated with MV (OR 1.03 [ 95% confidence interval (CI) 1.00, 1.05], p = 0.04; OR 2.10 [95% CI 1.03, 4.60], p = 0.04; OR 1.04 [ 95% CI 1.01, 1.07], p = 0.02); OR 0.80 [95% CI 0.66, 0.94], p = 0.02 and OR 0.65 [95% CI 0.52, 0.79, p

Clostridium botulinum and Clostridium tetani are Gram-positive, spore-forming, and anaerobic bacteria that produce the most potent neurotoxins, botulinum toxin (BoNT) and tetanus toxin (TeNT), responsible for flaccid and spastic paralysis, respectively. The main habitat of these toxigenic bacteria is the environment (soil, sediments, cadavers, decayed plants, intestinal content of healthy carrier animals). C. botulinum can grow and produce BoNT in food, leading to food-borne botulism, and in some circumstances, C. botulinum can colonize the intestinal tract and induce infant botulism or adult intestinal toxemia botulism. More rarely, C. botulinum colonizes wounds, whereas tetanus is always a result of wound contamination by C. tetani. The synthesis of neurotoxins is strictly regulated by complex regulatory networks. The highest levels of neurotoxins are produced at the end of the exponential growth and in the early stationary growth phase. Both microorganisms, except C. botulinum E, share an alternative sigma factor, BotR and TetR, respectively, the genes of which are located upstream of the neurotoxin genes. These factors are essential for neurotoxin gene expression. C. botulinum and C. tetani share also a two-component system (TCS) that negatively regulates neurotoxin synthesis, but each microorganism uses additional distinct sets of TCSs. Neurotoxin synthesis is interlocked with the general metabolism, and CodY, a master regulator of metabolism in Gram-positive bacteria, is involved in both clostridial species. The environmental and nutritional factors controlling neurotoxin synthesis are still poorly understood. The transition from amino acid to peptide metabolism seems to be an important factor. Moreover, a small non-coding RNA in C. tetani, and quorum-sensing systems in C. botulinum and possibly in C. tetani, also control toxin synthesis. However, both species use also distinct regulatory pathways; this reflects the adaptation of C. botulinum and C. tetani to different ecological niches. [ABSTRACT FROM AUTHOR]

Tetanus is a vaccine-preventable neuromuscular disease with a high mortality rate. The incidence of tetanus in developed countries has significantly declined due to preventive vaccination measures, but the potential for long-term complications and mortality from this disease remains high in the unvaccinated population. There are only a few individual case reports of tetanus in the pediatric population in the United States. We present a case of suspected tetanus in a 10-year-old unvaccinated child in Central Texas who sustained multiple cardiovascular and pulmonary complications during a 1-month hospitalization course. This case highlights the importance of pediatric immunization for prevention of this potentially fatal disease process and its long-term complications. Physicians should maintain a high clinical suspicion for tetanus infection in unvaccinated children to prevent delay in necessary treatment. [ABSTRACT FROM AUTHOR]

Tetanus is extremely rare in developed countries. We report the first documented case of tetanus in the province of Alberta since 2016: a farmer that developed trismus, shoulder stiffness, and fevers eight days following orthopedic surgery. Tetanus immunoglobulin elicited rapid recovery. We highlight risk factors, pathogenesis, epidemiology, and diagnostic challenges. [ABSTRACT FROM AUTHOR]

Tetanus is a life-threatening neurologic disease caused by an infectious agent, Clostridium Tetani, and is one of the infectious diseases that is preventable by vaccination. Tetanus is characterized by painful muscle spasm and increased muscle tone, that is caused by tetanospasmin, a powerful toxin elaborated by Clostridium tetani. Due to worldwide regular vaccination, this disease rarely seen recently. The case we present here is a patient admitted in Shohadaye khalije Fars hospital with severe generalized tetanus due to incomplete vaccination.

Tetanus is a neurologic non-transmissible disease (often fatal) of humans and other animals with a worldwide occurrence. Clostridium tetani is the spore producing bacillus which causes the bacterial disease. In deep penetrating wounds the spores germinate and produce a toxin called tetanospasmin. The main characteristic sign of tetanus is a spastic paralysis. A diagnosis is usually based on the clinical signs because the detection in the wound and the cultivation of C. tetani is very difficult. Between animal species there is considerable variability in the susceptibility to the bacillus. The most sensitive animal species to the neurotoxin are horses. Sheep and cattle are less sensitive and tetanus in these animal species are less common. Tetanus in cats and dogs are rare and dogs are less sensitive than cats. Clinically two forms of tetanus have been recognized, i. e. localized and generalized. The available treatment is not specific because the toxin in neuronal cells cannot be accessed by antitoxin antibodies. The aim of the therapy is to: neutralise the unbound neurotoxin, inhibit C. tetani growth in the wound, and provide supportive care to mitigate the effects of the neurotoxin. The treatment is difficult with an unclear prognosis.

Non-neonatal tetanus is an acute, specific, toxic disease in patients over 28 days of age, characterized by continuous rigidity and paroxysmal spasms of the skeletal muscles throughout the body caused by the intrusion of Clostridium tetani through skin or mucosal membrane into the body and reproducing in anaerobic environments to produce exotoxins. The mortality rate of severe patients is close to 100% without medical intervention. Even with aggressive comprehensive treatment, the global mortality rate remains at 30%-50%, making it a potentially fatal disease. In order to standardize the diagnosis, treatment and prevention of non-neonatal tetanus, based on "Regulation for Diagnosis and Treatment of Non-neonatal Tetanus (2019 Edition)", experts have revised this regulation according to clinical practice and recent research progress in this field to guide medical institutions in the prevention and control of non-neonatal tetanus. This article interprets the key points and basis for updating the 2024 edition regulation to guide clinical implementation and application.

Tetanus, caused by Clostridium tetani , is a significant health problem, particularly in regions lacking proper vaccination coverage against tetanus. Although it is usually associated with an identifiable wound, diagnosing tetanus without a visible entry site can be difficult. We report a 68-year-old man who was diagnosed with tetanus without any visible wounds or recent trauma. He had classic clinical features, including trismus, risus sardonicus, muscle spasms, and a positive spatula test, which all justified the clinical diagnosis of tetanus. The patient was treated with intramuscular injection of 3,000 IU of tetanus immune globulin, intravenous metronidazole, and muscle relaxants. He improved significantly after 10 days. He was discharged with follow-up and vaccination instructions. This case emphasizes the importance of maintaining a high index of suspicion for tetanus, even without any wound. It also reveals the effectiveness of clinical diagnostic methods and appropriate, timely treatment in rescuing the patient., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Sivagurunathan et al.)